Ligand-induced effects on pyruvate dehydrogenase kinase isoform 2

The Journal of Biological Chemistry
Yasuaki HiromasaThomas E Roche

Abstract

Tryptophan fluorescence was used to analyze binding of ligands to human pyruvate dehydrogenase isoform 2 (PDHK2) and to demonstrate effects of ligand binding on distal structure of PDHK2 that is required for binding to the inner lipoyl domain (L2) of the dihydrolipoyl acetyltransferase. Ligand-altered binding of PDHK2 to L2 and effects of specific ligands on PDHK2 oligomeric state were characterized by analytical ultracentrifugation. ATP, ADP, and pyruvate markedly quenched the tryptophan fluorescence of PDHK2 and gave maximum quenching/L0.5 estimates: approximately 53%/3 microM for ATP; approximately 49%/15 microM for ADP; and approximately 71%/approximately 590 microM for pyruvate. The conversion of Trp-383 to phenylalanine completely removed ATP- and ADP-induced quenching and > or = 80% of the absolute decrease in fluorescence due to pyruvate. The W383F-PDHK2 mutant retained high catalytic activity. Pyruvate, added after ADP, quenched Trp fluorescence with an L0.5 of 3.4 microM pyruvate, > or = 150-fold lower concentration than needed with pyruvate alone. ADP-enhanced binding of pyruvate was maintained with W383F-PDHK2. Binding of PDHK2 dimer to L2 is enhanced when L2 are housed in oligomeric structures, including the glutat...Continue Reading

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Citations

Jul 17, 2008·Biochemistry·Alla KlyuyevaKirill M Popov
Mar 30, 2017·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Shao-Lin ZhangKin Yip Tam
Jan 24, 2020·Cellular and Molecular Life Sciences : CMLS·Russel J ReiterLuiz G de Almeida Chuffa
Jan 31, 2021·Biophysical Journal·Bodhi A Jelinek, Michael A Moxley
May 24, 2021·Biochimica Et Biophysica Acta. Reviews on Cancer·Saleha AnwarMd Imtaiyaz Hassan

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