Mutational analysis reveals only one catalytic histidine in neutral endopeptidase ("enkephalinase").
Aside from serving as zinc ligands, kinetic data has implicated one or more additional histidines as catalytic residues in neutral endopeptidase ("enkephalinase") action. One of these histidines has previously been identified as histidine 704 (Bateman et al., J. Biol. Chem., 265:8365-8368, 1990). In order to determine whether a second histidine is involved in catalysis each of these residues not previously changed have been converted to glutamine by site directed mutagenesis. The resultant recombinant enzymes possess full catalytic activity indicating that histidine 704 is the only catalytic histidine in the enzyme.
Kim
Y A
YA
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235.
Shriver
B
B
Hersh
L B
LB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D006639:Histidine; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015260:Neprilysin / Q000235:genetics* / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D006639:Histidine; D015260:Neprilysin; D004047:Diethyl Pyrocarbonate