Primary structure of three peptides at the catalytic and allosteric sites of the fructose-1,6-bisphosphate-activated pyruvate kinase from Escherichia coli.
Three peptides containing 6-pyridoxyllysine have been isolated from the tryptic digest of the allosteric fructose-1,6-bisphosphate-dependent pyruvate kinase from Escherichia coli, which had been almost completely inactivated with pyridoxal 5'-phosphate. The labelled peptides have been sequenced. The comparison of their sequences with the primary structure of the cat muscle pyruvate kinase allowed to state that peptide I fits the region spanning residues 423-438 (53% identity), peptide II corresponds to residues 442-457 (44% identity) and peptide III encompasses residues 342-368 (70% identity). These findings are discussed in connection with our previous results on the involvement of the three peptides in the catalytic and regulatory properties of the enzyme (Valentini, G., Speranza, M.L., Iadarola, P., Ferri, G. & Malcovati, M. (1988) Biol. Chem. Hoppe-Seyler 369, 1219-1226) and in connection with their location in the three-dimensional structure of the cat muscle pyruvate kinase (Muirhead, H., Clayden, D.A., Lorimer, C.G., Fothergill-Gilmore, L.A., Schiltz, E. & Schmitt, W. (1986) EMBO J. 5, 475-481).
Speranza
M L
ML
Dipartimento di Biochimica, Università di Pavia.
Valentini
G
G
Iadarola
P
P
Stoppini
M
M
Malcovati
M
M
Ferri
G
G
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002415:Cats; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology*; D005635:Fructosediphosphates / Q000302:isolation & purification / Q000378:metabolism*; D006598:Hexosediphosphates / Q000378:metabolism*; D006868:Hydrolysis; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis*; D011487:Protein Conformation; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D011770:Pyruvate Kinase / Q000378:metabolism*; D051381:Rats; D014357:Trypsin
D000596:Amino Acids; D005635:Fructosediphosphates; D006598:Hexosediphosphates; D007202:Indicators and Reagents; D010455:Peptides; D011732:Pyridoxal Phosphate; D011770:Pyruvate Kinase; D014357:Trypsin; C029063:fructose-1,6-diphosphate