Rationalization of allosteric pathway in Thermus sp. GH5 methylglyoxal synthase
In: BMB Reports, Jg. 45 (2012), Heft 12, S. 748-753
Online
academicJournal
Zugriff:
A sequence of 10 amino acids at the C-terminus region ofmethylglyoxal synthase from Escherichia coli (EMGS) providesan arginine, which plays a crucial role in forming a salt bridgewith a proximal aspartate residue in the neighboring subunit,consequently transferring the allosteric signal between subunits.In order to verify the role of arginine, the gene encoding MGSfrom a thermophile species, Thermus sp. GH5 (TMGS) lackingthis arginine was cloned with an additional 30 bp sequence atthe 3´-end and then expressed in form of a fusion TMGS with a10 residual segment at the C-terminus (TMGS+). The resultingrecombinant enzyme showed a significant increase in cooperativitytowards phosphate, reflected by a change in the Hillcoefficient (nH) from 1.5 to 1.99. Experiments including sitedirected mutagenesis for Asp-10 in TMGS and TMGS+, twodimentional structural survey, fluorescence and irreversiblethermoinactivation were carried out to confirm this pathway.
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Rationalization of allosteric pathway in Thermus sp. GH5 methylglyoxal synthase
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Autor/in / Beteiligte Person: | Khajeh, Khosro |
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Zeitschrift: | BMB Reports, Jg. 45 (2012), Heft 12, S. 748-753 |
Veröffentlichung: | Korean Society for Biochemistry and Molecular Biology, 2012 |
Medientyp: | academicJournal |
ISSN: | 1976-6696 (print) ; 1976-670X (print) |
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