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The proteome of the bacterium Mycoplasma pneumoniae : Comparing predicted open reading frames to identified gene products
Author(s) -
Ueberle Barbara,
Frank Rainer,
Herrmann Richard
Publication year - 2002
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/1615-9861(200206)2:6<754::aid-prot754>3.0.co;2-2
Subject(s) - orfs , proteome , biology , open reading frame , isoelectric point , biotinylation , peptide mass fingerprinting , gene , proteomics , affinity chromatography , microbiology and biotechnology , peptide sequence , biochemistry , genetics , enzyme
An existing proteome map of the bacterium Mycoplasma pneumoniae comprising proteins from 224 genes was extended to 305 genes. This corresponds to about 44% of the 688 proposed genome sequence derived open reading frames (ORFs). The newly assigned gene products were enriched, separated by one‐dimensional or two‐dimensional (2‐D) gel electrophoresis and identified by mass spectrometry. The enrichment procedures included differential centrifugation, anion and cation exchange chromatography, affinity chromatography with heparin as a ligand and isolation of biotinylated proteins by binding to immobilized streptavidin. A comparative analysis of the identified proteins from 305 genes with the as yet unverified 383 ORFs concerning isoelectric point, molecular weight and number of transmembrane segments revealed that proteins with more than three predicted transmembrane segments and an isoelectric point above 10.5 are most likely not to be separated by 2‐D gel electrophoresis. The mutual benefits of genomics and proteomics were shown by the identification of a todate unannotated 128 amino acid long protein.