Biology Commons^™

A Study Of Inactive Enzyme-Homologues: The Biochemical And Biological Function Of Ecm14 In Saccharomyces Cerevisiae, Robert Christian Mcdonald

Master's Theses

Like most major enzyme families, the M14 family of metallocarboxypeptidases (MCPs) contains several pseudoenzymes predicted to lack enzyme activity and with unknown molecular function. The genome of the yeast Saccharomyces cerevisiae encodes only one member of the M14 MCP family, a pseudoenzyme named Ecm14 proposed to function in the extracellular matrix. Ecm14 is found throughout ascomycete fungi, with a group of related pseudoenzymes found in basidiomycetes. Although the prodomain of Ecm14 can be cleaved in vivo and in vitro by endopeptidases, suggesting an activation mechanism, no activity has been detected using standard carboxypeptidase substrates.

In order to examine the function …

Expression And Characterization Of Ecm14, A Metallocarboxypeptidase From Saccharomyces Cerevisiae, Matthew James Schott

Master's Theses

Metallocarboxypeptidases are found in most organisms and function in the digestion and maturation of proteins. Ecm14 is a putative metallocarboxypeptidase found in the yeast Saccharomyces cerevisiae vacuole. The function of Ecm14 as an enzyme has been unclear due to the presence of active site amino acids not typically found in metallocarboxypeptidases, suggesting either no enzymatic activity or a unique mechanism. In order to investigate the enzymatic mechanism of Ecm14, expression of histidine-tagged Ecm14 protein was attempted in human HEK293T cell culture, S. cerevisiae, and baculovirus expression systems. No expression was detected in HEK293T cells in preliminary experiments. Expression in the …