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Acrylamide gel electrophoresis of hydrophobic proteins: Gas vacuole protein
Author(s) -
Simon Robert D.
Publication year - 1980
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.1150010310
Subject(s) - vacuole , chemistry , chromatography , polyacrylamide gel electrophoresis , sodium dodecyl sulfate , halobacterium , gel electrophoresis , biochemistry , anabaena , acrylamide , molecular weight size marker , cucumis , gel electrophoresis of proteins , cyanobacteria , bacteria , biology , cytoplasm , polymer , monomer , botany , organic chemistry , enzyme , membrane , bacteriorhodopsin , genetics
The hydrophobic protein comprising the gas vacuoles of a variety of procaryotic microorganisms is insoluble in detergents ( e.g. sodium dodecyl sulfate) and cannot be electrophoresed in detergent‐containing systems. The protein is solubilized by mixtures of phenol‐acetic acid‐urea (PAU) and the development of a slab gel system containing PAU which successfully separates the gas vacuole protein is described. In the system, phenol is polymerized directly into the gel, and the high background staining which results can be minimized by the proper choice of gel thickness, acrylamide/Bis ratio, and polymerization conditions. There is a direct relationship between the electrophoretic mobility and molecular weight of protein standards in the PAU system; however, several proteins including histones H 2 , H 3 , H 4 and bovine serum albumin have a faster rate of gration than expected. It has been possible to separate and identify the gas vacuole protein from Halobacterium halobium, Halobacterium salinarium strain 5, and the two species of cyanobacteria, Anabaena flos‐aquae and Microcystis aeruginosa . Mobility of the gas vacuole proteins in the four species are consistent with molecular weights of 16 800, 16 800, 14 700 and 15 700 respectively. Gels produced using the PAU system can be successfully fluorographed.