Characterization of porins isolated from the outer membrane of Vibrio damsela.
A protein oligomer with an approximate molecular weight of its 37-kDa monomer form was purified from the cell envelope fraction of Vibrio damsela cells. This oligomer exhibited strong porin activity when reconstituted into proteoliposomes with phosphatidyl choline. The functional properties for the 37-kDa protein suggest that it is a nonspecific or general porin, with an apparent pore size of 1.6 nm. This porin allows penetration of a variety of hydrophilic solutes according to their molecular mass. After electroelution, the oligomer was partially dissociated into monomers, whereas treatment with EDTA did not affect its dissociation. The monomers of the 37-kDa protein were not active in the reconstitution assay. The effect of culture media on the composition of the outer membrane protein of V. damsela was examined. Only one outer membrane protein with an apparent molecular weight of 37 kDa (37-kDa protein) was formed in cells grown in 3% NaCl-BHI broth and in 3% NaCl-nutrient broth with the addition of 2% glucose. Three outer membrane proteins, with apparent molecular weights of 37 kDa, 40 kDa, and 46 kDa, were produced in cells grown in 3% NaCl-nutrient broth. An additional outer membrane protein with an apparent molecular weight of 44 kDa (44-kDa protein) was found in cells grown in 3% NaCl-nutrient broth with the addition of 2% maltose. This protein was found to exhibit specificity to maltose derivatives. The results obtained in this study confirm the porin-like character of discussed proteins and give a basis for advanced study of those proteins.
Nitzan
Y
Y
Health Sciences Research Center, Department of Life Sciences, Bar-Ilan University, Ramat Gan 52900, Israel.
Gribun
A
A
Pechatnikov
I
I
D015153:Blotting, Western; D003470:Culture Media / Q000737:chemistry; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D008320:Maltose / Q000494:pharmacology; D016366:Open Reading Frames; D018272:Porins / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification; D012965:Sodium Chloride / Q000494:pharmacology; D014733:Vibrio / Q000737:chemistry* / Q000254:growth & development
D003470:Culture Media; D018272:Porins; D012965:Sodium Chloride; D008320:Maltose; D004492:Edetic Acid