Single Molecule Studies of Membrane Proteins on Glass Substrates using Atomic Force Microscopy
Creators: name: Chada, Nagaraju
name: Sigdel, Krishna
name: Matin, Tina
name: Sanganna Gari, Raghavendar Reddy
name: Mao, Chunfeng
name: Randall, Linda
name: King, Gavin
Publisher: name: Zenodo
Year: 2014
Subjects: subject: Biological Physics
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Biochemistry
subject: Atomic force microscope imaging
subject: AFM
subject: Protein Translocation
subject: Bacteriorhodopsin bR
subject: Escherichia coli
subject: Halobacterium salinarum
subject: SecA ATPase motor protein
subject: Single Molecule
subject: Membrane Proteins
subject: Glass Chemical Treatment
subject: Glass Etching
subject: Glass Micro-machining
subject: Protein-protein interactions
subject: Surface roughness of Glass
subject: Surface roughness of Mica
subject: Surface roughness of Lipid bilayer
subject: Midwest Single Molecule Workshop
subject: Lipid Membrane
subject: Purple Membrane
subject: Transmembrane protein
subject: SecYEG
subject: Sec Pathway
subject: Motor Proteins

description: 1) Method for for integration of high resolution biological AFM with other powerful optical techniques 2) Straight-forward cleaning procedure for treatment of glass for Microscopy and Micromachining applications 3) Molecular high resolution imaging of bacteriorhodopsin and Sec translocase on glass supports 4) Direct observation of protein-protein interactions 5) Atomic Force Microscopy measurements of surface roughness of Glass, Mica, Lipid and comparison of several glass chemical treatments for Microscopy <strong>READ THE PEER-REVIEWED PUBLICATION HERE</strong> <strong>ASSOCIATED PEER-REVIEWED PUBLICATION </strong> <strong>Abstract</strong> Since its invention in the mid-1980s, the atomic force microscope (AFM) has become a valuable complementary<br> tool for studying membrane proteins in near-native environments. Historically, mica is the most common<br> substrate utilized for biological AFM. Glass being amorphous, transparent, and optically homogeneous has its<br> own set of advantages over mica and has the potential to broaden the overlap of AFM with techniques that require<br> high quality non-birefringent optical access. The use of silanized glass as an AFM substrates has been reported as<br> a means to fine tune surface chemistry. However, such coatings usually require hours of additional preparation<br> time and can lead to increased surface roughness. In this work, we present a simple technique for preparing<br> borosilicate glass as a substrate for two membrane protein systems: non-crystalline translocons (SecYEG) of the<br> general secretary system from E. coli, and bacteriorhodopsin (BR) from H. salinarum. For both these membrane<br> proteins, quantitative comparisons of the measured protein structures on glass versus mica substrates show<br> agreement. An additional advantage of glass is that lipid coverage is rapid (&lt; 1 minute) and complete (occupying<br> the entire surface). A goal is to study the bacterial export system using recently developed precision measurement<br> techniques such as ultra-stable AFM.
description: {"references": ["Chada, N., Sigdel, K., Gari, R. et al. Glass is a Viable Substrate for Precision Force Microscopy of Membrane Proteins. Sci Rep 5, 12550 (2015). https://doi.org/10.1038/srep12550", "Chada, N., Sigdel, K., Matin, T., Sanganna Gari, R.R., Mao, C., Randall, L. and King, G., 2013. Glass is a viable substrate for atomic force microscopy of membrane proteins. Bulletin of the American Physical Society, 58. http://meetings.aps.org/Meeting/PSF13/Event/206952", "Chada, N., Sigdel, K.P., Matin, T.R., Gari, R.R.S., Mao, C., Randall, L.L. and King, G.M., 2014. Glass is a viable substrate for atomic force microscopy of membrane proteins. Biophysical Journal, 106(2), p.458a., https://www.cell.com/biophysj/pdf/S0006-3495(13)03853-8.pdf", "Chada, N., Sigdel, K.P., Reddy, R., Gari, S., Matin, T.R., Mao, C., Marsh, B., Randall, L.L. and King, G.M., 2015. Glass: A Multi-Platform Specimen Supporting Substrate for Precision Single Molecule Studies of Membrane Proteins. Biophysical Journal, 108(2), p.170a., https://www.cell.com/biophysj/pdf/S0006-3495(14)02145-6.pdf", "N. Chada, Watching biological nanomotors at work: insights from single-molecule studies, (2017), Thesis, University of Missouri-Columbia, Missouri, USA, Link: , https://mospace.umsystem.edu/xmlui/bitstream/handle/10355/63612/research.pdf.pdf?sequence=1&amp;isAllowed=y", "Chada , Nagaraju . Growth and Characterization of Transition Metal Oxides for Chemical Sensor Applications: Setting up Initiated Hot Wire Chemical Vapor Deposition System. (2010)."]}