bluuebunny/crossref_metadata_2025 huggingface dataset:
The present and future of China's National Science and Technology Library
Authors: Meng Liansheng , Quan Liu Yan
Published: July 2005

Purpose

The purpose of this case study is to explore the construction pattern and issues involved with the development of the National Science and Technology Library (NSTL) in China.

Design/methodology/approach

This study was conducted through an analytical evaluation of the historical development of scientific and technological information sharing among libraries and information centers and issues around construction of the NSTL.

Findings

Since the establishment of its network service system at the end of 2000, the NSTL has made rapid progress in constructing a sci‐tech information resource sharing system and offering web‐based information services to the patrons of scientific and technical communities all over China.

Originality/value

This paper is the first exploration that brings to the forefront the foundational background of the NSTL, the characteristics of its organizational structure, the functions of its service, and the tentative plan to increase its accessibility and usability. Symptomatic of the function of the NSTL constructing pattern will hopefully shed light not only to the reformation of the Chinese sci‐tech information management system, but also to the development of scientific digital libraries in the world.
Comparing polarized perspectives: librarians' professional skills and development
Authors: Gosine‐Boodoo Meerabai , McNish Mark
Published: July 2005

Purpose

The main aim of the research conducted was to identify whether the particular country environment of today's professional librarian impacts upon his/her skills capabilities as well as upon his/her access to opportunities for continued development.

Design/methodology/approach

Data was sourced by means of a survey administered to professional librarians mainly from the developing Caribbean Region and the developed North American countries. Firstly, the satisfaction level of librarians with regard to six recommended skills and characteristics was measured. These areas are detailed as follows: communication; training; information technology (IT); managerial; commitment and subject knowledge/profiling. Secondly, professional development opportunities were measured via skills, services and attitudes, key areas also recommended for staff development and training.

Findings

Findings demonstrated that librarians' perceptions showed noteworthy similarities and fewer differences than expected between the respective country categories. Both perspectives reflected similar zones of “skills insecurity”.

Research limitations/implications

The greatest challenge faced by the researchers was to source a sample size that could yield meaningful results in relation to the hypotheses.

Originality/value

A comparison of librarians from a geo‐economic position (i.e. in relation to geographic location and world economic order) appears to have been largely unexplored. The study is intended as well to encourage key players from both country contexts to unite for the enhancement of librarians' professional development.
Serving the “Boomer” generation and beyond
Authors: Long Sarah Ann
Published: July 2005

Purpose

To focus on the importance of providing appropriate and specialized services for the senior population.

Design/methodology/approach

Body of knowledge compiled by author from current news, professional sources and personal topical files.

Findings

Libraries in the US have a wide variety of creative programs and services for senior‐aged patrons, as well as handicapped users.

Originality/value

The column stresses the importance of libraries to study and plan for the challenges of serving a rapidly growing senior population.
The role of information technology in supply‐chain relationships: does partner criticality matter?
Authors: Kim Daekwan , Tamer Cavusgil S.
Published: July 2005

Purpose

There have been contradictory findings in the literature regarding the impact of information technology (IT) on firm productivity. While the debate known as the “IT paradox” still endures, there has been little empirical research to clarify why or when IT offers benefits to the owning firms. This study attempts to fill this gap in the literature by investigating when IT contributes to firm performance.

Design/methodology/approach

Using survey data, this study explores how IT adoption by a firm, in the context of supply chain communication systems, influences its market performance. Of special interest are improvements in both the firm's own and its partner's coordination activities. Criticality of a firm's channel partner is used as a moderating variable.

Findings

Results suggest that a firm's own coordination mediates the influence of IT adoption on market performance only when the partner being coordinated is critical to its success. If the partner is not critical to the success of the firm, the benefits of IT adoption can be materialized only through enhancements in the coordination activities of the partner.

Originality/value

The study results clarify why the assumed benefits of IT have been inconsistent in the context of supply chain relationships.
Critical factors affecting intermediary web site adoption: understanding how to extend e‐participation
Authors: Harrison Tina , Waite Kathryn
Published: July 2005

Purpose

To provide an investigation of e‐commerce development via an examination of the forces shaping web site development among intermediaries in an extended supply chain.

Design/methodology/approach

A two‐stage research design combining qualitative and quantitative methods. Unstructured interviews conducted in the spirit of phenomenology elicited a range of critical incidents of web site development which were further examined via a quantitative survey of intermediaries to test for relationships between critical incidents and web site adoption.

Findings

Adopter groups were identified which showed statistically significant differences in terms of the critical incidents driving web site development as well as differences in terms of key company characteristics. The timing of web site adoption was also found to affect the subsequent use of the technology, with early adopters making more advanced use.

Research limitations/implications

Limitations associated with the use of retrospective data and respondents’ abilities to recall events, although attempts were made to minimise these through external validation.

Practical implications

Provides useful insights for providers of financial services in understanding how to progress the adoption of web site technology by intermediaries, suggesting the development of networks of relationships involving IT suppliers rather than simply focusing on relationships with preferred intermediaries.

Originality/value

Addresses a research gap in terms of business‐to‐business e‐commerce and offers practical guidance on how to widen participation in the financial services supply chain.
Inter‐organisational collaboration for the digital economy
Authors: Houldsworth Elizabeth , Alexander Gillian
Published: July 2005

Purpose

This paper was intended to explore the concept of learning in networks using recent literature from this emerging field. In order to do so, it takes as a case study a pan‐European e‐learning project, which required collaborative working and learning within a distributed team.

Design/methodology/approach

The approach has been qualitative and exploratory. Data have been collected via interviews with project partners and participant observation of the steering group meetings. Analysis has similarly been qualitative in nature.

Findings

Analysis suggests that there are links with earlier literature on inter‐organisational learning networks, particularly around culture, management style and leadership in virtual collaboration. Interestingly, even supposed “experts” in the field of e‐learning technologies showed a preference to work face‐to‐face.

Research limitations/implications

The research to date has been exploratory and small‐scale. More work is required to test the findings in a wider context.

Practical implications

The findings are believed to have considerable practical value, both for the European Union, as the funding body, but also for one's own practice, and that of others as they attempt to facilitate learning and collaborating in networks.

Originality/value

The paper adds value as it is one of only a small number to approach networked learning from a non‐experimental standpoint. It is also original in its application of the frameworks around network learning and learning in networks, as advanced by Knight in 2002 and Knight and Pye in 2002.
An empirical framework developed for selecting B2B e‐business models: the case of Australian agribusiness firms
Authors: Ng Eric
Published: July 2005

Purpose

To develop a preliminary framework for Australian agribusiness organisations seeking to select business‐to‐business (B2B) e‐business models.

Design/methodology/approach

A literature review was conducted on topics related to strategic decision making and B2B e‐business models. Particularly, factors influencing the strategic decision on the choice of e‐business models to be selected were examined. The review aims to provide an initial framework for the research study to be conducted. The research was conducted in two stages: depth interviews (stage one) and case studies (stage two).

Findings

Provides information on factors (both internal and external) influencing the choice of e‐business models and also insight into the current practices of Australian agribusiness in relation to the selection process of B2B e‐business models. A framework was developed to assist agribusiness organisations to make decisions on the selection of the most appropriate e‐business models.

Research limitations/implications

This study is exploratory in nature and thus the findings cannot be generalised to the population at large. Further conclusive explanatory research is required for generalisation and the guidelines developed in this study could be replicated and tested in other agribusiness sectors or in other industries.

Practical implications

Managers are able to consider and examine the relevance of the guidelines and criteria developed, and determine the essential factors that require consideration during their selection process. The guidelines can also assist managers to determine the level of resources, technological infrastructure and knowledge and understanding of e‐business models required.

Originality/value

This paper brings together two disciplines – strategic decision making and development of e‐business models – that have not been combined prior to this study. The research findings have contributed to the development of existing theory in these two areas. The research also offers insights into the selection of e‐business models within the agribusiness industry that traditionally lagged behind in e‐business. The development of the framework and guidelines has assisted managers in their selection of e‐business models and given them an appreciation of what others in the industry are doing.
A decision‐support system for business‐to‐business marketing
Authors: Noori Behrooz , Hossein Salimi Mohammad
Published: July 2005

Purpose

The main purpose of this paper is to review the related literature and propose a new decision‐support‐system (DSS) framework for marketing in the business‐to‐business (B2B) arena based on customer‐relationship management (CRM) and knowledge‐driven marketing to help related‐field graduate students and marketing managers.

Design/methodology/approach

Reviews a range of the most important works published between 1966 and 2004 in order to demonstrate both practical and theoretical aspects. The main method of this research is analytical and conceptual and the approach to this subject was to investigate the gap between marketing DSSs and analytical CRM.

Findings

Provides information about a customized marketing DSS in a B2B context, indicates related literature and frameworks and, finally, tests the ideas with a case study.

Practical implications

Outcomes and applications are identified for developing new activities in improving marketing decision making and marketing planning based on customer orientation and customer satisfaction.

Originality/value

Despite such interdependencies, the research in the fields of DSSs and CRM solutions has not adequately considered the integration of such systems. The novel contribution of this paper lies in integrating marketing DSSs and CRM with regard to knowledge‐driven marketing in B2B marketing, in both theoretical and practical aspects.
Why doesn't marketing use the corporate data warehouse? The role of trust and quality in adoption of data‐warehousing technology for CRM applications
Authors: Cobb Payton Fay , Zahay Debra
Published: July 2005

Purpose

This paper aims to investigate organizational factors to explain why a corporate data warehouse (CDW) was not used by marketing to the extent that it was expected to be used for CRM and other marketing purposes.

Design/methodology/approach

A case study of a single health‐care payor organization is used in this study.

Findings

Reveals the three primary implementation factors related to marketing's lack of trust in the data, low perceived data quality and perception of marketing needs not being met. Practically, the unique data needs of marketing should be considered in the implementation of a CDW and its interface.

Originality/value

This is the first study of its kind to take the needs of marketing users into consideration.
Unintended responses to a traditional purchasing performance metric
Authors: Emiliani M.L. , Stec D.J. , Grasso L.P.
Published: July 2005

Purpose

To describe the tactics that buyers often use to avoid unfavorable purchase price variance (PPV) and identify alternate approaches that will improve purchasing performance and also help achieve company objectives.

Design/methodology/approach

Descriptive: presents for the first time 12 dysfunctional tactics used by buyers of industrial goods use to avoid unfavorable PPV.

Findings

The tactics are shown to increase costs rather than decrease costs and lead to organizational dysfunction. Findings are broadly applicable to large corporations that use legacy software systems or newer enterprise requirement planning (ERP) software systems to track purchasing costs and transactions, and also have a strong management focus on price‐based purchasing performance.

Research limitations/implications

Findings are limited to organizations that measure the success of purchasing and supply management activities using price‐based metrics.

Practical implications

Should propel managers to identify alternative metrics or processes for managing purchasing performance, reduce system‐wide costs, and improve day‐to‐day work in purchasing organizations.

Originality/value

This paper will be helpful to academics researching operational or behavioral aspects of purchasing, practitioners managing supply chains, auditors assessing the integrity of material cost reporting and management controls, and persons concerned about ethics in business.
Emerging internet‐enabled auction mechanisms in supply chain
Authors: Nair Anand
Published: July 2005

Purpose

The purpose of this paper is to present the prevailing auction and bidding mechanisms in supply chain management. The paper aims to critically examine the implications of these auction mechanisms using the example of the transportation sector.

Design/methodology/approach

A framework is presented to highlight the role of internet and information systems in the transportation sector. The paper discusses the complexities associated with embracing these internet‐enabled auction mechanisms and presents some research propositions.

Findings

In its present state of affairs an electronic marketplace would become nothing but a virtual place where the competing firms would be indulged in price wars to maintain their presence. A prolonged spur of activities in this direction would lead to a stalemate where the winners are losers.

Practical implications

In the long run it is important that the decisions associated with electronic auctions must be based on more than bargain‐basement price tags. The business intangibles focused on developing a strong buyer‐supplier relationship, will determine the quality of the purchase contract.

Research implications

The propositions presented here could provide initial impetus for future empirical research that gleans insights regarding the link between the use of electronic auction mechanisms and the strength of buyer‐supplier relationships.

Originality/value

Provides a comprehensive review and some potential research propositions that could enhance our understanding of supply chain management.
Network as a business environment: experiences from software industry
Authors: Kulmala Harri I. , Uusi‐Rauva Erkki
Published: July 2005

Purpose

The aim of the paper is to describe the differences between networked and non‐networked firms' methods intended to increase profitability, methods for intended growth, implemented and intended openness development, and experiences of networking.

Design/methodology/approach

The research approach is empirical and descriptive. Data were gathered by mail survey from Finnish software producers.

Findings

Networked firms intended to improve profitability by boosting resource efficiency, while non‐networked firms concentrated on price increases. No difference occurred in firms' perceptions of their profitability. Networked firms intended to grow by mergers and acquisitions and by cooperation with partners. Non‐networked firms were eager to grow by developing new software products. Networked firms intended to increase information sharing more and were more open in sharing information concerning firms' technical competencies, cost, and commitments to other organizations. Lack of marketing resources was the most important reason for firms to join a network.

Research limitations/implications

The research setting (how the firms are classified in groups, for example) could benefit from deeper analysis of how network characteristics are connected and which variables best estimate and describe the degree of networking of a firm. A larger sample of firms should be covered to achieve generalizable results. Systematic comparison of industries would also provide essential information on network dynamics.

Originality/value

Networked firms had grown more than non‐networked firms but no connection between profitability perceptions and networking was obtained. In software industry, growth may produce networks but networking seems to support growth.
Electronic shipping documentation in China's international supply chains
Authors: Mei Zhiliang , Dinwoodie John
Published: July 2005

Purpose

Before China's opening and reform policy transformed foreign trade, personal relationships characterised mainly domestic trade. Burgeoning international supply chains (ISC) require efficient document transfers to accompany large cargo shipments, but China's users of bills of lading (BOLs) increasingly experience delays or fraud. The purpose of this study is to investigate the potential for an internet‐based, third‐party (IBTP) internet service provider (ISP) offering electronic (e)BOLs and shipping documentation services.

Design/methodology/approach

An exploratory attitude survey of BOL users compared trader, middlemen and shipping group perceptions’ of current problems and potential for eBOLS.

Findings

Accuracy, security and speed subsumed cost considerations for BOL users. Major concerns were delays, typing errors and excessive paperwork. EBOLs proffer speedier documentation, better management information and savings on postage. Concerns for secure data with legal integrity signify mistrust, favouring electronic data interchange (EDI).

Research implications

This work studied BOL users in Shanghai. Future comparisons of how trust develops as other ISCs adopt eBOLS should identify groups with particular interests and potential industrial partners, evaluate user‐costs, savings and ability to surmount existing problems.

Practical implications

More traders trusted eBOLS but extensive training is required through state support or partnerships with internet‐based, third‐party internet service providers (IBTPIS) to establish infrastructure, knowledge, confidence and trust amongst potential local users. EBOL data is considered insecure. Its legal status must be clarified.

Originality/value

This paper alerts participants in China's ISCs that trust suffuses trade. It quantifies user valuations of electronic systems that accelerate shipping documentation processes and advocates training of shipper and middlemen groups to dispel mistrust.
Looking beyond value‐based pricing of beef in North America
Authors: Tronstad Russell , Unterschultz James
Published: July 2005

Purpose

Quality traits desired by consumers may not be adequately captured by beef industry standards associated with grid or value‐based pricing alone. Aims to demonstrate this shortcoming by examining strategies of selected companies in North America at the four supply chain levels of cow‐calf genetics, feedlot feeding, processing, and retailing that have been proactive in producing desirable beef attributes efficiently to better meet consumer beef demand.

Design/methodology/approach

The vertical alliance between Ralphs retailing, Sunland Beef processing, and a handful of feedlots using narrowly defined beef genetics are examined to illustrate how consumer market research and coordination throughout the supply chain may address many shortcomings associated with current value‐based pricing of beef criteria.

Findings

Better information sharing and coordination between seedstock and retail industries could help assure that consumer preferences of beef palatability and consistency are met while meeting high production efficiency standards.

Practical implications

Cow‐calf, feedlot, and packing industries need to better track and manage information flows of genetic‐management paths from consumer to seedstock producer in order for the beef industry to be more competitive.

Originality/value

Experiences of our case companies suggest that the beef industry will need to look beyond the North American grid or value‐based pricing of beef in order to maintain or improve market share with competing pork and poultry sectors.
Benchmarking in health services
Authors: Guven‐Uslu Pinar
Published: July 2005

Purpose

To investigate implementation of benchmarking (BM) in three hospital trusts by adapting receptive context of change model according to BM principles of EFQM business model.

Design/methodology/approach

Perceptions of managers, clinicians and finance personnel towards implementation of BM are compared. Both qualitative and quantitative data are collected in three hospitals and results are cross compared to analyse both organisational and professional perceptions.

Findings

The paper concludes that there are limits to the rapid or broad implementation of BM principles in health services. It argues that the patients and their expectations are not referred in BM. It also suggests that local implementation programmes should be used for BM health services.

Research limitations/implications

Future research could use data from other hospitals and apply same/similar framework in order to create knowledge representing the general attitude in hospitals and between professionals towards BM in health services.

Practical implications

Both academics and professionals working in the field of health services management might find the paper useful especially in managing different professional groups' attitude in managing change in large organisations.

Originality/value

The originality of the paper is the fact that it attempts to report on both professional and organisational approaches in implementation of BM. It also attempts to report on not only organisational factors but also individual and external factors of change for BM.
Performance measurement for green supply chain management
Authors: Hervani Aref A. , Helms Marilyn M. , Sarkis Joseph
Published: July 2005

Purpose

To introduce and provide an overview of the various issues related to environmental (green) supply chain management performance measurement.

Design/methodology/approach

The work relies on experiences, case studies and other literature related to performance measurement in environmental supply chains. It seeks to integrate works in supply chain management, environmental management, and performance management into one framework. A systems framework forms the discussion outline with a focus on controls/pressures, inputs, tools, and outputs as major categories for evaluation and review.

Findings

Provides an integrative framework for study, design and evaluation of green supply chain management performance tools. The findings also identify a number of issues that need to still be addressed.

Research limitations/implications

We have only one design of the issues in which numerous categorizations could be provided. There is limited research in this area and new and current models/developments can provide additional insight. Implications of the work is that these gaps exist and that significantly more work needs to be completed in this field.

Practical implications

A very useful source of information for practitioners that seek to implement these systems within and between organizations. Also, the paper provides numerous areas which researchers could complete additional research and develop research agendas.

Originality/value

This paper provides some of the very first insights into development of a green supply chain management performance measurement system. Typically performance measurement systems are internally and business focused, we expand on these issues by considering inter‐organizational and environmental issues within a business context.
Employee benefits of multi‐owner accounting firms: groundwork for benchmarking
Authors: McGaughey Ronald , Puleo Victor , Casey K. Michael
Published: July 2005

Purpose

The purpose of this research paper is to provide practitioners and researchers with guidance and ideas for benchmarking employee benefits in companies providing professional services. The research addressed employee benefits in multi‐owner accounting firms.

Design/methodology/approach

Data from a survey of a large number of multi‐owner accounting firms (CPA firms) were analyzed to examine professional employee benefits and to look at the relationship between firm size and benefits offered.

Findings

An analysis of survey results suggested that larger firms offer better benefits than smaller firms. Larger firms tend also to be more profitable. Various employee benefit metrics were examined.

Research limitations/implications

The survey was limited to accounting firms in the United States, so the findings may have limited value for researchers and practitioners in other countries.

Practical implications

The better benefits offered by larger accounting firms may allow them to attract better personnel, possibly accounting for their greater profitability. If this is indeed true, then a good benefit package may well be a key success factor for accounting firms, and possibly for other professional services. Firms seeking to improve their competitive position may, therefore, find it advantageous to benchmark their professional employee benefits against the benefit packages of larger more profitable competitors.

Originality/value

This paper examines professional employee benefits in multi‐owner accounting firms and identifies metrics that could be useful to practitioners in benchmarking those benefits. The metrics identified and other findings may provide practitioners with ideas for benchmarking benefits in other professional service organizations.
Sustainability: necessity for a prosperous society
Authors: Fokkema Jacob , Jansen Leo , Mulder Karel
Published: July 2005

Purpose

To present the challenge of sustainable development, the way in which technology can address that challenge and the task of engineering education to train engineers for it.

Design/methodology/approach

The paper describes briefly the history of the environmental and sustainability discourse in The Netherlands, as a densely populated country. It argues that technology should play a major role in SD, but that technological innovation is not enough. Technological systems renewal is a transdisciplinary activity involving relevant stakeholders and disciplines. “Needs” is the basic starting‐point to innovate new systems of provision. The paper reviews relevant literature regarding future orientation of technology development. Based on it, goals for training of engineers are developed.

Findings

The engineer has to meet a threefold challenge: providing new creative approaches on the one hand, and setting up and executing R&D programs that produce results, on the other; cooperating with other disciplines and lay stakeholders, on the one hand, and guarding disciplinary quality, on the other; bridging moralism and strategic pragmatism.

Research limitations/implications

The paper is an introduction, i.e. it sketches the issues without dealing with them in detail.

Practical implications

The paper draws in broad lines a road‐map for the future of engineering education and sustainable development. The paper is a useful source for those engineering institutions that are formulating a strategy to introduce sustainable development.

Originality/value

The paper goes beyond environmental engineering, not by just adding social and economic issues, but by developing an integrated framework for academic training of engineers.
Learning about environmental issues
Authors: Lundholm Cecilia
Published: July 2005

Purpose

To present results from a research project on postgraduate and undergraduate students' learning about environmental issues in education.

Design/methodology/approach

Three cases were carried out with civil engineering students, biology students and postgraduate students. Discussions in classroom were tape‐recorded, as well as discussions while working with assignments, and interviews were carried out.

Findings

Shows how differently environmental issues can be interpreted, i.e. scientifically, existentially and politically, and the way values and emotions become an aspect of the learning process and reveal the students' difficulties in differentiating between values and descriptions of phenomena. This is analysed and explained in relation to the students' various projects that come into conflict in the educational setting.

Practical implications

The paper can be of use to those who are engaged in environmental education and raises questions regarding the content of today's environmental education.

Originality/value

Gives an insight into students' learning processes and experiences in environmental education and the difficulties they can experience when studying such a subject.
Introduction of sustainable development in engineers' curricula
Authors: Lourdel N. , Gondran N. , Laforest V. , Brodhag C.
Published: July 2005

Purpose

Owing to its complexity, sustainable development cannot be simply integrated as a supplementary course within the engineers' curricula. The first point of this paper aims to focalise on how to reflect pedagogically. After dealing with these questions, a tool that can evaluate the student's understanding of sustainable development concepts will be presented.

Design/methodology/approach

The analysis of a student's sustainability comprehension, based on cognitive maps, has been developed. The students are asked to write and connect by arrows all the terms that they associate with the concept of sustainable development. The assessment of the aforementioned cognitive maps is based on an approach via semantic category.

Findings

This study shows that the students' perception of sustainable development before the training seems mainly focalised on environmental and economical aspects. After the SD course, an increase in the number of words quoted is noted for each category (social and cultural aspects; the stakeholders, the principles of sustainable development and the allusions to complexity, temporal and spatial dimensions). Their vision seems richer and wider. The training seemed useful to help the students who did not associate sustainable development with diverse dimension to improve this perception.

Research limitations/implications

To reduce the length of the elaboration of the maps, there is no preliminary training for the construction of maps. To simplify this elaboration, only one type of arrows is used to connect words. To minimise the time of analysis of the maps, the relevance of the relations made between the words is not verified. Besides, the classification of words within the semantic categories implies a certain level of subjectivity.

Practical implications

This cognitive map method can be a useful tool to improve learning in quantitative terms but also in qualitative terms. Identifying knowledge gaps and misunderstood ideas allows the improvement in the training.

Originality/value

This study presents a new method that can be used to evaluate the impact of training sessions on students. Another advantage is to analyse how the students' knowledge is interconnected. This seems particularly interesting because the study of this transdisciplinary concept also necessitates an integrated vision.
A novel system for continuous protein refolding and on‐line capture by expanded bed adsorption
Authors: Ferré Henrik , Ruffet Emmanuel , Nielsen Lise‐Lotte B. , Nissen Mogens Holst , Hobley Timothy J. , Thomas Owen R.T. , Buus Søren
Published: July 2005

A novel two‐step protein refolding strategy has been developed, where continuous renaturation‐bydilution is followed by direct capture on an expanded bed adsorption (EBA) column. The performance of the overall process was tested on a N‐terminally tagged version of human β2‐microglobulin (HAT‐hβ2m) both at analytical, small, and preparative scale. In a single scalable operation, extracted and denatured inclusion body proteins from Escherichia coli were continuously diluted into refolding buffer, using a short pipe reactor, allowing for a defined retention and refolding time, and then fed directly to an EBA column, where the protein was captured, washed, and finally eluted as soluble folded protein. Not only was the eluted protein in a correctly folded state, the purity of the HAThβ2m was increased from 34% to 94%, and the product was concentrated sevenfold. The yield of the overall process was 45%, and the product loss was primarily a consequence of the refolding reaction rather than the EBA step. Full biological activity of HAT‐hβ2m was demonstrated after removal of the HAT‐tag. In contrast to batch refolding, a continuous refolding strategy allows the conditions to be controlled and maintained throughout the process, irrespective of the batch size; i.e., it is readily scalable. Furthermore, the procedure is fast and tolerant toward aggregate formation, a common complication of in vitro protein refolding. In conclusion, this system represents a novel approach to small and preparative scale protein refolding, which should be applicable to many other proteins.
The intrinsically disordered C‐terminal domain of the measles virus nucleoprotein interacts with the C‐terminal domain of the phosphoprotein via two distinct sites and remains predominantly unfolded
Authors: Bourhis Jean‐Marie , Receveur‐Bréchot Véronique , Oglesbee Michael , Zhang Xinsheng , Buccellato Matthew , Darbon Hervé , Canard Bruno , Finet Stéphanie , Longhi Sonia
Published: July 2005

Measles virus is a negative‐sense, single‐stranded RNA virus within theMononegavirales order,which includes several human pathogens, including rabies, Ebola, Nipah, and Hendra viruses. Themeasles virus nucleoprotein consists of a structured N‐terminal domain, and of an intrinsically disordered C‐terminal domain, NTAIL (aa 401–525), which undergoes induced folding in the presence of the C‐terminal domain (XD, aa 459–507) of the viral phosphoprotein. With in NTAIL, an α‐helical molecular recognition element (α‐MoRE, aa 488–499) involved in binding to P and in induced folding was identified and then observed in the crystal structure of XD. Using small‐angle X‐ray scattering, we have derived a low‐resolution structural model of the complex between XD and NTAIL, which shows that most of NTAIL remains disordered in the complex despite P‐induced folding within the α‐MoRE. The model consists of an extended shape accommodating the multiple conformations adopted by the disordered N‐terminal region of NTAIL, and of a bulky globular region, corresponding to XD and to the C terminus of NTAIL (aa 486–525). Using surface plasmon resonance, circular dichroism, fluorescence spectroscopy, and heteronuclear magnetic resonance, we show that NTAIL has an additional site (aa 517–525) involved in binding to XD but not in the unstructured‐to‐structured transition. This work provides evidence that intrinsically disordered domains can establish complex interactions with their partners, and can contact them through multiple sites that do not all necessarily gain regular secondary structure.
Remodeling protein complexes: Insights from the AAA+ unfoldase ClpX and Mu transposase
Authors: Burton Briana M. , Baker Tania A.
Published: July 2005

Multiprotein complexes in the cell are dynamic entities that are constantly undergoing changes in subunit composition and conformation to carry out their functions. The protein–DNA complex that promotes recombination of the bacteriophage Mu is a prime example of a complex that must undergo specific changes to carry out its function. The Clp/Hsp100 family of AAA+ ATPases plays a critical role in mediating such changes. The Clp/Hsp100 unfolding enzymes have been extensively studied for the roles they play in protein degradation. However, degradation is not the only fate for proteins that come in contact with the ATP‐dependent unfolding enzymes. The Clp/Hsp100 enzymes induce structural changes in their substrates. These structural changes, which we refer to as “remodeling,” ultimately change the biological activity of the substrate. These biological changes include activation, inactivation (not associated with degradation), and relocation within the cell. Analysis of the interaction between Escherichia coli ClpX unfoldase and the Mu recombination complex, has provided molecular insight into the mechanisms of protein remodeling. We discuss the key mechanistic features of the remodeling reactions promoted by ClpX and possible implications of these findings for other biological reactions.
Interdomain side‐chain interactions in human γD crystallin influencing folding and stability
Authors: Flaugh Shannon L. , Kosinski‐Collins Melissa S. , King Jonathan
Published: July 2005

Human γD crystallin (HγD‐Crys) is a two domain, β‐sheet eye lens protein that must remain soluble throughout life for lens transparency. Single amino acid substitutions of HγD‐Crys are associated with juvenile‐onset cataracts. Features of the interface between the two domains conserved among γ‐crystallins are a central six‐residue hydrophobic cluster, and two pairs of interacting residues flanking the cluster. In HγD‐Crys these pairs are Gln54/Gln143 and Arg79/Met147. We previously reported contributions of the hydrophobic cluster residues to protein stability. In this study alanine substitutions of the flanking residue pairs were constructed and analyzed. Equilibrium unfolding/refolding experiments at 37°C revealed a plateau in the unfolding/refolding transitions, suggesting population of a partially folded intermediate with a folded C‐terminal domain (C‐td) and unfolded N‐terminal domain (N‐td). The N‐td was destabilized by substituting residues from both domains. In contrast, the C‐td was not significantly affected by substitutions of either domain. Refolding rates of the N‐td were significantly decreased for mutants of either domain. In contrast, refolding rates of the C‐td were similar to wild type for mutants of either domain. Therefore, domain interface residues of the folded C‐td probably nucleate refolding of the N‐td. We suggest that these residues stabilize the native state by shielding the central hydrophobic cluster from solvent. Glutamine and methionine side chains are among the residues covalently damaged in aged and cataractous lenses. Such damage may generate partially unfolded, aggregation‐ prone conformations of HγD‐Crys that could be significant in cataract.
Refractive index‐based determination of detergent concentration and its application to the study of membrane proteins
Authors: Strop Pavel , Brunger Axel T.
Published: July 2005

The concentration of detergent in membrane protein preparations can have a critical role on protein stability, function, and the potential for crystallization. Unfortunately, dialysis or protein concentration can lead to an unknown amount of detergent in the final membrane protein preparations. Here we present a method for the determination of detergent concentration based on refractive index of the detergent solution. This method was applied to quantitate the amount of detergent remaining in solution after concentration in various concentrators. We found that the ability of the tested detergents to pass through the molecular weight cutoff membrane correlates well with detergent micelle size. Therefore, the micelle size can be used as a rough guide to estimate the retention of a given detergent in various molecular weight cutoff concentrators. The refractive index method is exceptionally informative when coupled with size exclusion chromatography and light scattering, and can be used to determine the oligomeric state of the membrane protein, the size of a protein‐associated micelle, as well as the amount and size of the unbound detergent micelle.
Three‐dimensional structure of the AAH26994.1 protein from Mus musculus, a putative eukaryotic Urm1
Authors: Singh Shanteri , Tonelli Marco , Tyler Robert C. , Bahrami Arash , Lee Min S. , Markley John L.
Published: July 2005

We have used NMR spectroscopy to determine the solution structure of protein AAH26994.1 from Mus musculus and propose that it represents the first three‐dimensional structure of a ubiquitin‐related modifier 1 (Urm1) protein. Amino acid sequence comparisons indicate that AAH26994.1 belongs to the Urm1 family of ubiquitin‐like modifier proteins. The best characterized member of this family has been shown to be involved in nutrient sensing, invasive growth, and budding in yeast. Proteins in this family have only a weak sequence similarity to ubiquitin, and the structure of AAH26994.1 showed a much closer resemblance to MoaD subunits of molybdopterin synthases (known structures are of three bacterial MoaD proteins with 14%–26% sequence identity to AAH26994.1). The structures of AAH26994.1 and the MoaD proteins each contain the signature ubiquitin secondary structure fold, but all differ from ubiquitin largely in regions outside of this fold. This structural similarity bolsters the hypothesis that ubiquitin and ubiquitin‐related proteins evolved from a protein‐based sulfide donor system of the molybdopterin synthase type.
Mutual paracrine effects of colorectal tumour cells and stromal cells: modulation of tumour and stromal cell differentiation and extracellular matrix component production in culture
Authors: Mukaratirwa Sydney , Koninkx Jos F. , Gruys Erik , Nederbragt Hubertus
Published: July 2005

Interactions of tumour and stromal cells influence tumour cell proliferation and differentiation, stromal cell phenotypic transdifferentiation and secretion of extracellular matrix (ECM) components. In this study, we established a monolayer and a three‐dimensional cell‐to‐cell interaction model between canine mammary stromal cells and human colonic carcinoma cell lines (Caco‐2 and HT‐29) to investigate mutual paracrine effects of tumour cells and stromal cells on (i) tumour cell differentiation, (ii) production of ECM components and (iii) phenotypic transdifferentiation of stromal cells. We showed that when Caco‐2 or HT‐29 cells are cultured in collagen gels, they form a few small solid cell clusters with no lumina, but when cocultured with stromal cells, the tumour cells formed glandular structures with central lumina. This fibroblast‐induced organization and differentiation of Caco‐2 cells (not HT‐29 cells) appeared to be mediated by transforming growth factor‐β (TGF‐β). Culturing of stromal cells, Caco‐2 cells or HT‐29 cells alone in both monolayers and gels resulted in weak tenascin‐C expression in stromal cells and HT‐29 cells and no expression in the Caco‐2 cells. Coculturing of stromal cells with tumour cells resulted in increased tenascin‐C expression in the stromal cells and HT‐29 cells and induced expression of tenascin‐C in the Caco‐2 cells. This induction and increased expression of tenascin‐C appeared to be mediated by TGF‐β. Culturing of stromal cells, Caco‐2 cells or HT‐29 cells alone on monolayers and in gels resulted in a weak expression of chondroitin sulfate (CS), chondroitin‐6‐sulfate (C‐6‐S) and versican in stromal cells and no expression in Caco‐2 and HT‐29 cells. Coculturing of stromal cells with tumour cells on monolayers and in gels resulted in increased CS, C‐6‐S and versican expression in stromal cells. This tumour cell‐induced expression of CS, C‐6‐S and versican appeared to be mediated by TGF‐β and platelet‐derived growth factor (PDGF). Coculturing of Caco‐2 and HT‐29 and stromal cells promoted the transdifferentiation of stromal cells into myofibroblasts, and this appeared to be mediated by TGF‐β. These results suggest that TGF‐β and PDGF are part of a paracrine system involved in stromal–epithelial cell interaction important in stromal cell differentiation and ECM component production.
Dietary copper supplements modulate aortic superoxide dismutase, nitric oxide and atherosclerosis
Authors: Lamb David J. , Tickner Michelle L. , Hourani Susanna M. O. , Ferns Gordon A. A.
Published: July 2005

The objective was to test the hypothesis that dietary copper inhibits atherosclerosis by inducing superoxide dismutase (SOD) and potentiating nitric oxide (NO). New Zealand White rabbits were fed either a cholesterol diet (n = 8) or a cholesterol diet containing 0.02% copper acetate (n = 8) for 13 weeks. We found that the intimal area was significantly smaller in the animals supplemented with copper (P < 0.005), although integrated plasma cholesterol levels were not significantly different. This was associated with a significant increase in aortic copper content (P < 0.05), SOD activity (P < 0.05) and Cu/Zn SOD mRNA (P < 0.05) and a significant decrease in nitrotyrosine content (P < 0.05). Furthermore, there was a positive correlation between aortic copper content and SOD activity (P < 0.005,R2 = 0.83) and a negative correlation between aortic superoxide dimutase activity and nitrotyrosine content (P < 0.005,R2 = 0.93). In organ bath experiments, the relaxation of precontracted carotid artery rings to calcium ionophore was greater in animals supplemented with copper. No difference in response to sodium nitroprusside was observed. These data suggest that in the cholesterol‐fed rabbit, copper supplements inhibit the progression of atherosclerosis by increasing SOD expression, thereby reducing the interaction of NO with superoxide, and hence potentiating NO‐mediated pathways that may protect against atherosclerosis.