6PRX

oxidized Human Branched Chain Aminotransferase mutant C318A

Summary for 6PRX

• Entry DOI: 10.2210/pdb6prx/pdb
• Descriptor: Branched-chain-amino-acid aminotransferase, mitochondrial, PYRIDOXAL-5'-PHOSPHATE (2 entities in total)
• Functional Keywords: oxidized, metabolic role, conformational change, transferase
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 2
• Total formula weight: 87200.42

Authors

Dong, M.,Herbert, D.,Gibbs, S. (deposition date: 2019-07-11, release date: 2020-01-22, Last modification date: 2023-11-15)

Primary citation

Herbert, D.,Gibbs, S.,Riddick, A.,Conway, M.,Dong, M.
Crystal structure of an oxidized mutant of human mitochondrial branched-chain aminotransferase.
Acta Crystallogr.,Sect.F, 76:14-19, 2020

PubMed Abstract: This study presents the crystal structure of a thiol variant of the human mitochondrial branched-chain aminotransferase protein. Human branched-chain aminotransferase (hBCAT) catalyzes the transamination of the branched-chain amino acids leucine, valine and isoleucine and α-ketoglutarate to their respective α-keto acids and glutamate. hBCAT activity is regulated by a CXXC center located approximately 10 Å from the active site. This redox-active center facilitates recycling between the reduced and oxidized states, representing hBCAT in its active and inactive forms, respectively. Site-directed mutagenesis of the redox sensor (Cys315) results in a significant loss of activity, with no loss of activity reported on the mutation of the resolving cysteine (Cys318), which allows the reversible formation of a disulfide bond between Cys315 and Cys318. The crystal structure of the oxidized form of the C318A variant was used to better understand the contributions of the individual cysteines and their oxidation states. The structure reveals the modified CXXC center in a conformation similar to that in the oxidized wild type, supporting the notion that its regulatory mechanism depends on switching the Cys315 side chain between active and inactive conformations. Moreover, the structure reveals conformational differences in the N-terminal and inter-domain region that may correlate with the inactivated state of the CXXC center.

PubMed: 31929181
DOI: 10.1107/S2053230X19016480

Experimental method

X-RAY DIFFRACTION (3.25 Å)