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Identification of the active site serine of hormone‐sensitive lipase by site‐directed mutagenesis
Author(s) -
Holm Cecilia,
Davis Richard C.,
Østerlund Torben,
Schotz Michael C.,
Fredrikson Gudrun
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)00403-x
Subject(s) - serine , pentapeptide repeat , lipase , site directed mutagenesis , esterase , active site , biochemistry , hormone sensitive lipase , serine hydrolase , monoacylglycerol lipase , mutagenesis , chemistry , biology , mutant , enzyme , peptide , gene , endocannabinoid system , receptor
The consensus pentapeptide GXSXG is found in virtually all lipases/esterases and generally contains the active site serine. The primary sequence of hormone‐sensitive lipase contains a single copy of this pentapeptide, surrounding Ser‐423. We have analyzed the catalytic role of Ser‐423 by site‐directed mutagenesis and expression of the mutant hormone‐sensitive lipase in COS cells. Substitution of Ser‐423 by several different amino acids resulted in the complete abolition of both lipase and esterase activity, whereas mutation of other conserved serine residues had no effect on the catalytic activity. These results strongly suggest that Ser‐423 is the active site serine of hormone‐sensitive lipase.