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Evidence that the acyl‐ O ‐esters are intermediates in the catalysis
Author(s) -
McCarthy Alun D.,
Grahame Hardie D.
Publication year - 1982
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(82)81330-6
Subject(s) - iodoacetamide , acyl carrier protein , enzyme , biochemistry , chemistry , acetyl coa , fatty acid synthase , acyl group , coenzyme a , fatty acid , atp synthase , stereochemistry , cysteine , biosynthesis , organic chemistry , group (periodic table) , reductase
The sequence acetyl‐CoA → acetyl‐ O ‐enzyme → acetyl‐ S ‐acyl carrier protein has for the first time been demonstrated directly with a multifunctional (mammalian) fatty acid synthase. This was achieved by blocking of the active‐site thiols of rabbit mammary fatty acid synthase with iodoacetamide. The modified enzyme was incubated with [ 14 C]acetyl‐CoA to form acetyl‐ O ‐enzyme, and acetyl‐CoA was removed rapidly by centrifuge desalting. We were then able to demonstrate transfer of the acetyl group from [ 14 C]acetyl‐ O ‐enzyme to the pantetheine thiol in a fragment of rabbit mammary fatty acid synthase containing the phosphopantetheine group, and to E. coli acyl carrier protein.