Copper metalloproteomics
Subtitle: using immobilized metal affinity chromatography, two-dimensional gel electrophoresis and mass spectrometry to search for hepatocellular proteins with copper-binding ability.
Published: 2004
Language: eng
Number of pages: 114
Description: Copper is an essential trace element that participates in diverse biochemical reactions in all aerobic organisms. We have outlined the make up of the human copper metalloproteome. HepG2 cytosolic and microsomal fractions were applied to a copper(II)-loaded immobilized metal affinity chromatography (IMAC) column. Bound proteins were eluted with 50 mM EDTA, separated by 2D-gel electrophoresis and identified by MALDI-QqTOF mass spectrometry. We identified 48 cytosolic proteins and 19 microsomal proteins with copper-binding ability. To our knowledge, this is the first effort to describe the composition of the copper metalloproteome. Members of the 2-Cys family of peroxiredoxins (Prxs) were identified in this study but were not previously known to bind copper. Results presented here indicate that Prx3, a 25 kDa mitochondrial protein with peroxidase activity, converts Cu(II) to Cu(I) and binds Cu(I) with a stoichiometry of 2 mol Cu(I) per monomer of protein. The physiological relevance of Prx3 copper-binding is unclear.
Notes: Adviser:  Bibudhendra Sarkar.

Thesis (M.Sc.)--University of Toronto, 2004.

Electronic version licensed for access by U. of T. users.

Source: Masters Abstracts International, Volume: 43-03, page: 0850.

MICR copy on microfiche (2 microfiches).