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PMID: 19081061 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Structural basis for inactivation of the human pyruvate dehydrogenase complex by phosphorylation: role of disordered phosphorylation loops.

Structure (London, England : 1993) ·Vol. 16 ·No. 12 ·2008-12-10 ·Pages 1849-59

Kato M, Wynn RM, Chuang JL, Tso SC, Machius M, Li J, Chuang DT

Abstract

We report the crystal structures of the phosporylated pyruvate dehydrogenase (E1p) component of the human pyruvate dehydrogenase complex (PDC). The complete phosphorylation at Ser264-alpha (site 1) of a variant E1p protein was achieved using robust pyruvate dehydrogenase kinase 4 free of the PDC core. We show that unlike its unmodified counterpart, the presence of a phosphoryl group at Ser264-alpha prevents the cofactor thiamine diphosphate-induced ordering of the two loops carrying the three phosphorylation sites. The disordering of these phosphorylation loops is caused by a previously unrecognized steric clash between the phosphoryl group at site 1 and a nearby Ser266-alpha, which nullifies a hydrogen-bonding network essential for maintaining the loop conformations. The disordered phosphorylation loops impede the binding of lipoyl domains of the PDC core to E1p, negating the reductive acetylation step. This results in the disruption of the substrate channeling in the PDC, leading to the inactivation of this catalytic machine.

MeSH Terms
Acetylation Alanine/metabolism Amino Acid Substitution Binding Sites/genetics Decarboxylation Humans Hydrogen Bonding Kinetics Models, Biological Models, Molecular Phosphorylation Protein Binding/genetics Protein Conformation Protein Kinases/metabolism Protein Structure, Tertiary/genetics Pyruvate Dehydrogenase (Lipoamide)/genetics,metabolism Pyruvate Dehydrogenase Complex/chemistry,genetics,metabolism Substrate Specificity/genetics Thiamine Pyrophosphate/genetics,metabolism
Chemicals
Pyruvate Dehydrogenase Complex Pyruvate Dehydrogenase (Lipoamide) Protein Kinases pyruvate dehydrogenase kinase 4 Alanine Thiamine Pyrophosphate
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Kato Masato
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390-9038, USA.
Wynn R Max
Chuang Jacinta L
Tso Shih-Chia
Machius Mischa
Li Jun
Chuang David T
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Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
0969-2126
DOI
10.1016/j.str.2008.10.010
Published
2008-12-10
Pages
1849-59
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC2849990
Subset
IM
Grants
NIDDK NIH HHS · R56 DK062306-06A1 · United States
NIDDK NIH HHS · R01 DK026758 · United States
NIDDK NIH HHS · DK62306 · United States
NIDDK NIH HHS · R01 DK062306-05 · United States
NIDDK NIH HHS · R56 DK062306 · United States
NIDDK NIH HHS · R01 DK062306 · United States
NIDDK NIH HHS · R01 DK026758-30 · United States
NIDDK NIH HHS · DK26758 · United States
External Links
PubMed source DOI: 10.1016/j.str.2008.10.010 PMC Full Text View journal details
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