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Directed mutagenesis of pig renal membrane dipeptidase His 219 is critical but the DHXXH motif is not essential for zinc binding or catalytic activity
Author(s) -
Keynan Shoshana,
Hooper Nigel M.,
Turner Anthony J.
Publication year - 1994
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(94)00637-7
Subject(s) - dipeptidase , site directed mutagenesis , chemistry , biochemistry , zinc , mutant , enzyme , mutagenesis , binding site , microbiology and biotechnology , biology , gene , organic chemistry
Pig renal membrane dipeptidase cDNA has been expressed in COS‐1 cells. Directed mutagenesis was used to investigate the roles of some conserved histidyl and aspartyl residues. Mutation of His 219 to Arg, Lys or Leu results in complete abolition of enzyme activity, although the mutants are expressed at the cell‐surface. Residues in a proposed motif (DHXDH; residues 269−273) for zinc binding have been mutated individually. Each retained activity comparable to that of the wild‐type, excluding an essential role for components of this motif. The zinc‐binding ligands in membrane dipeptidase therefore represent a novel domain for a metallopeptidase with His 219 being one candidate.