Identification of a cross-linked double-peptide from the catalytic site of the Ca(2+)-ATPase of sarcoplasmic reticulum formed by the Ca(2+)- and pH-dependent reaction with ATP gamma P-imidazolidate.
The Ca(2+)-ATPase from sarcoplasmic reticulum can be inhibited by the Ca(2+)- and pH-dependent reaction with ATP gamma P-imidazolidate. The chemically and monofunctionally activated inhibitor introduces an intramolecular cross-link between two neighbouring peptides of the active site. This can be followed by the reduced mobility of the ATPase upon SDS-PAGE analysis which becomes even more pronounced after limited trypsinolysis. After cleavage of the cross-linked ATPase molecule by cyanogen bromide and separation of the peptides a double-peptide can be detected which upon sequencing can be identified as part of the phosphorylation and the nucleotide binding site, respectively.
Gutowski-Eckel
Z
Z
Institut für Biophysikalische Chemie und Biochemie, Johann-Wolfgang-Goethe Universität, Frankfurt/Main, Germany.
Mann
K
K
Bäumert
H G
HG
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D003432:Cross-Linking Reagents; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D010455:Peptides / Q000737:chemistry; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*
D003432:Cross-Linking Reagents; D007093:Imidazoles; D010455:Peptides; C057152:ATP gammaP-imidazolidate; D000255:Adenosine Triphosphate; D000252:Calcium-Transporting ATPases; D002118:Calcium