The activity of a thermostable lipoyl synthase from Sulfolobus solfataricus with a synthetic octanoyl substrate
The activity of a thermostable lipoyl synthase from Sulfolobus solfataricus with a synthetic octanoyl substrate
The protein lipoyl synthase (LipA) is essential for lipoic acid biosynthesis via sulfur insertions into a protein-bound octanoyl group. We have developed an in vitro assay for LipA using a synthetic tetrapeptide Substrate, containing an N-epsilon-octanoyl lysine residue, corresponding in sequence to the lipoyl binding domain of the E2 subunit of pyruvate dehydrogenase. A putative LipA from the hypothermophilic archaea Sulfolobus solfataricus was expressed in Escherichia coli and purified, and the activity was measured using this novel assay. The optimal temperature for the S. solfataricus LipA-dependent formation of the lipoyl group was found to be 60 degrees C.
44-49
Bryant, P.
b25c6534-2db1-4843-8b14-fb72613b2be6
Kriek, M.
e01ddb12-38a7-4733-9e13-c0e5d9670cca
Wood, R.J.
6694b71e-a4ec-49fd-b2a4-644a7534f9f0
Roach, P.L.
ca94060c-4443-482b-af3e-979243488ba9
2006
Bryant, P.
b25c6534-2db1-4843-8b14-fb72613b2be6
Kriek, M.
e01ddb12-38a7-4733-9e13-c0e5d9670cca
Wood, R.J.
6694b71e-a4ec-49fd-b2a4-644a7534f9f0
Roach, P.L.
ca94060c-4443-482b-af3e-979243488ba9
Bryant, P., Kriek, M., Wood, R.J. and Roach, P.L.
(2006)
The activity of a thermostable lipoyl synthase from Sulfolobus solfataricus with a synthetic octanoyl substrate.
Analytical Biochemistry, 351 (1), .
(doi:10.1016/j.ab.2006.01.023).
Abstract
The protein lipoyl synthase (LipA) is essential for lipoic acid biosynthesis via sulfur insertions into a protein-bound octanoyl group. We have developed an in vitro assay for LipA using a synthetic tetrapeptide Substrate, containing an N-epsilon-octanoyl lysine residue, corresponding in sequence to the lipoyl binding domain of the E2 subunit of pyruvate dehydrogenase. A putative LipA from the hypothermophilic archaea Sulfolobus solfataricus was expressed in Escherichia coli and purified, and the activity was measured using this novel assay. The optimal temperature for the S. solfataricus LipA-dependent formation of the lipoyl group was found to be 60 degrees C.
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Published date: 2006
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Local EPrints ID: 44399
URI: http://eprints.soton.ac.uk/id/eprint/44399
ISSN: 0003-2697
PURE UUID: 80351c92-b910-4e7e-b6d5-a8cc3518de30
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Date deposited: 06 Mar 2007
Last modified: 15 Mar 2024 09:03
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Author:
P. Bryant
Author:
M. Kriek
Author:
R.J. Wood
Author:
P.L. Roach
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