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Nitroprusside stimulates the cysteine‐specific mono(ADP‐ribosylation) of glyceraldehyde‐3‐phosphate dehydrogenase from human erythrocytes
Author(s) -
Kots Alexander Ya.,
Skurat Alexander V.,
Sergienko Edward A.,
Bulargina Tamara V.,
Severin Eugene S.
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80153-8
Subject(s) - nad+ kinase , biochemistry , glyceraldehyde 3 phosphate dehydrogenase , adp ribosylation , microbiology and biotechnology , adenylate kinase , chemistry , dithiothreitol , dehydrogenase , cysteine , glycerol 3 phosphate dehydrogenase , hydroxylamine , monoclonal antibody , immunoprecipitation , enzyme , biology , antibody , gene , immunology
In human erythrocyte membranes incubated with [adenylate‐ 32 P]NAD the 36 kDa protein is predominantly labeled. The labeling is greatly stimulated by nitroprusside in the presence of dithiothreitol. We have purified the 36 k Da protein and identified this modification as crysteine‐specific mono(ADP‐ribosylation) because: (i) labeling occured only when [ 32 P]NAD was replaced by adenine [U‐ 14 C]NAD, but not by [carbonyl‐ 14 C]NAD; (ii) treatment of the prelabeled protein with snake venom phosphodiesterase led to releasing 5′‐[ 32 P]AMP; (iii) the bond between the protein and the nucleotide was hydrolyzed by HgCl 2 , but was resistant to hydroxylamine. The 36 kDa protein reacted on Western blots with two different monoclonal antibodies (MAbs) against glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) and was immunoprecipitated by both MAbs.