Proteome of the Escherichia coli envelope and technological challenges in membrane proteome analysis.
The envelope of Escherichia coli is a complex organelle composed of the outer membrane, periplasm-peptidoglycan layer and cytoplasmic membrane. Each compartment has a unique complement of proteins, the proteome. Determining the proteome of the envelope is essential for developing an in silico bacterial model, for determining cellular responses to environmental alterations, for determining the function of proteins encoded by genes of unknown function and for development and testing of new experimental technologies such as mass spectrometric methods for identifying and quantifying hydrophobic proteins. The availability of complete genomic information has led several groups to develop computer algorithms to predict the proteome of each part of the envelope by searching the genome for leader sequences, beta-sheet motifs and stretches of alpha-helical hydrophobic amino acids. In addition, published experimental data has been mined directly and by machine learning approaches. In this review we examine the somewhat confusing available literature and relate published experimental data to the most recent gene annotation of E. coli to describe the predicted and experimental proteome of each compartment. The problem of characterizing integral versus membrane-associated proteins is discussed. The E. coli envelope proteome provides an excellent test bed for developing mass spectrometric techniques for identifying hydrophobic proteins that have generally been refractory to analysis. We describe the gel based and solution based proteome analysis approaches along with protein cleavage and proteolysis methods that investigators are taking to tackle this difficult problem.
Weiner
Joel H
JH
Membrane Protein Research Group and The Institute for Biomolecular Design, University of Alberta, Canada. joel.weiner@ualberta.ca
Li
Liang
L
D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D029968:Escherichia coli Proteins / Q000032:analysis; D020869:Gene Expression Profiling; D020411:Oligonucleotide Array Sequence Analysis; D019897:Periplasm / Q000737:chemistry; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods* / Q000639:trends
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D020543:Proteome