Assignment of function to histidines 260 and 298 by engineering the E1 component of the Escherichia coli 2-oxoglutarate dehydrogenase complex; Substitutions that lead to acceptance of substrates lacking the 5-carboxyl group

Document Type

Article

Publication Date

9-6-2011

Abstract

The first component (E1o) of the Escherichia coli 2-oxoglutarate dehydrogenase complex (OGDHc) was engineered to accept substrates lacking the 5-carboxylate group by subjecting H260 and H298 to saturation mutagenesis. Apparently, H260 is required for substrate recognition, but H298 could be replaced with hydrophobic residues of similar molecular volume. To interrogate whether the second component would allow synthesis of acyl-coenzyme A derivatives, hybrid complexes consisting of recombinant components of OGDHc (o) and pyruvate dehydrogenase (p) enzymes were constructed, suggesting that a different component is the "gatekeeper" for specificity for these two multienzyme complexes in bacteria, E1p for pyruvate but E2o for 2-oxoglutarate. © 2011 American Chemical Society.

Identifier

80052213490 (Scopus)

Publication Title

Biochemistry

External Full Text Location

https://doi.org/10.1021/bi200936n

e-ISSN

15204995

ISSN

00062960

PubMed ID

21809826

First Page

7705

Last Page

7709

Issue

35

Volume

50

Grant

R01GM050380

Fund Ref

National Institute of General Medical Sciences

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