Pseudomonas aeruginosa cytotoxin-binding protein in rabbit erythrocyte membranes : an oligomer of 28 kDa with similarity to transmembrane channel proteins

• Published in: European journal of biochemistry Vol. 217; no. 3; pp. 1123 - 1128
• Main Authors: LUTZ, F, MOHR, M, GRIMMIG, M, LEIDOLF, R, LINDER, D
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell 01.11.1993
• Subjects: Amino Acid Sequence; Animals; Aquaporin 1; Aquaporins; Bacteriology; Biological and medical sciences; Blood Group Antigens; Erythrocyte Membrane - metabolism; Fundamental and applied biological sciences. Psychology; Humans; Ion Channels - metabolism; Leukocidins - metabolism; Membrane Proteins - chemistry; Membrane Proteins - metabolism; Microbiology; Molecular Sequence Data; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; Pseudomonas aeruginosa; Pseudomonas aeruginosa - metabolism; Rabbits; Sequence Homology, Amino Acid; Pseudomonadales; Transmembrane protein; Toxin; Plasma membrane; Red blood cell; Bacteria; Pseudomonadaceae; Molecular interaction; Pseudomonas aeruginosa; In vitro
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1993.tb18345.x

Rabbit erythrocyte membrane glycosylated 28-kDa protein was investigated in the membrane-bound 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, 125I-cytotoxin binds with high affinity to water-channel 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.