Open AccessPseudomonas aeruginosa cytotoxin‐binding protein in rabbit erythrocyte membranes
Author(s) -
LUTZ Frieder,
MOHR Manuela,
GRIMMIG Magdalene,
LEIDOLF Regina,
LINDER Dietmar
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb18345.x
Subject(s) - trypsin , biochemistry , membrane , peptide , western blot , binding protein , microbiology and biotechnology , protein g , nitrocellulose , chemistry , biology , enzyme , antibody , gene , immunology
Rabbit erythrocyte membrane glycosylated 28‐kDa protein was investigated in the membranebound as well as in the soluble state on an example of a Pseudomonas aeruginosa cytotoxin‐binding component. When membranes were treated with trypsin/ N ‐glycosidase F, a 13.5‐kDa‐binding active peptide residue is obtained as revealed by a ligand‐blot technique after separation by SDS/PAGE under reducing conditions and electrophoretic transfer to nitrocellulose. Target‐size analysis of intact membranes by radiation inactivation using 2–450 kGy gave a value of 29, 40 and 60 kDa for the binding‐protein structure. This suggests that the native form of the binding peptide is associated as an oligomer. As seen in ligand‐blot technique, 125 I‐cytotoxin binds with high affinity to waterchannel integral protein CHIP28 from human erythrocyte membranes. The 20 N‐terminal amino acids of the deglycosylated rabbit cytotoxin‐binding protein show high similarity to transmembrane channel‐like proteins.