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Two‐dimensional map of Haemophilus influenzae following protein enrichment by heparin chromatography
Author(s) -
Fountoulakis Michael,
Langen Hanno,
Evers Stefan,
Gray Chris,
Takács Béla
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150180728
Subject(s) - haemophilus influenzae , chemistry , chromatography , matrix assisted laser desorption/ionization , gel electrophoresis , electrophoresis , affinity chromatography , polyacrylamide gel electrophoresis , bacteria , biochemistry , mass spectrometry , biology , desorption , genetics , enzyme , organic chemistry , adsorption , antibiotics
Two‐dimensional gel electrophoresis separates several hundred protein molecules in one single experiment and is efficiently used to study the products expressed by different genomes. Low‐copy‐number gene products are invisible on a stained two‐dimensional map and must be enriched such that sufficient amounts are present for visualization and identification. We investigated the enrichment of proteins of the bacterium Haemophilus influenzae by chromatography on immobilized heparin which has affinity for growth and protein biosynthesis factors. Total soluble proteins of the microorganism were fractionated on Heparin‐Actigel which resulted in enrichment of approximately 160 proteins. The eluates, representing about 40% of the applied proteins, were analyzed by two‐dimensional gel electrophoresis and the protein spots were characterized by amino acid composition analysis and matrix‐assisted laser desorption ionization mass spectrometry. The proteins enriched by chromatography on the heparin gel were not exclusively low‐copy‐number gene products and they did not exclusively belong to one single class of proteins. The proteins that bound to the heparin gel are indicated in a two‐dimensional protein map which includes more than 110 newly identified proteins.