Article

From proteins to proteomes: large scale protein identification by two-dimensional electrophoresis and amino acid analysis.

Macquarie University Centre for Analytical Biotechnology, Macquarie University, Sydney, NSW, Australia.
Bio/Technolgy 02/1996; 14(1):61-5. pp.61-5
Source: PubMed

ABSTRACT Separation and identification of proteins by two-dimensional (2-D) electrophoresis can be used for protein-based gene expression analysis. In this report single protein spots, from polyvinylidene difluoride blots of micropreparative E. coli 2-D gels, were rapidly and economically identified by matching their amino acid composition, estimated pI and molecular weight against all E. coli entries in the SWISS-PROT database. Thirty proteins from an E. coli 2-D map were analyzed and identities assigned. Three of the proteins were unknown. By protein sequencing analysis, 20 of the 27 proteins were correctly identified. Importantly, correct identifications showed unambiguous "correct" score patterns. While incorrect protein identifications also showed distinctive score patterns, indicating that protein must be identified by other means. These techniques allow large-scale screening of the protein complement of simple organisms, or tissues in normal and disease states. The computer program described here is accessible via the World Wide Web at URL address (http:@expasy.hcuge.ch/).

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Keywords

27 proteins
 
amino acid composition
 
computer program
 
correct identifications
 
disease states
 
distinctive score patterns
 
identities
 
incorrect protein identifications
 
large-scale screening
 
molecular weight
 
normal
 
polyvinylidene difluoride blots
 
protein-based gene expression analysis
 
proteins
 
report single protein spots
 
score patterns
 
simple organisms
 
SWISS-PROT database
 
URL address
 
World Wide Web
 

M R Wilkins