Crystal structure of the binary complex of pig muscle phosphoglycerate kinase and its substrate 3-phospho-D-glycerate

K. Harlos, M. Vas, C. F. Blake

Research output: Contribution to journalArticle

115 Citations (Scopus)

Abstract

Pig muscle phosphoglycerate kinase has been crystallized from polyethyleneglycol in the presence of its substrate 3-phospho-D-glycerate (3- PG) and the structure has been determined at 2.0 Å resolution. The structure was solved using the known structure of the substrate-free horse muscle enzyme and has been refined to a crystallographic R-factor of 21.5%. 3- Phospho-D-glycerate is bound to the N-domain of the enzyme through a network of hydrogen bonds to a cluster of basic amino acid residues and by electrostatic interactions between the negatively charged phosphate and these basic protein side chains. This binding site is in good agreement with earlier proposals [Banks et al., Nature (London) 279:773-777, 1979]. The phosphate oxygen atoms are hydrogen bonded to His-62, Arg-65, Arg-122, and Arg-170. The 2-hydroxyl group, which defines the D-isomer of 3PG, is hydrogen bonded to Asp-23 and Asn-25. The carboxyl group of 3-PG points away from the N-domain towards the C-domain and is hydrogen bonded via a water molecule to main chain nitrogen atoms of helix-14. The present structure of the 3-PG- bound pig muscle enzyme is compared with the structure of the substrate-free horse enzyme. Major changes include an ordering of helix-13 and a domain movement, which brings the N-domain closer to the ATP-binding C-domain. This domain movement consists of a 7.7° rotation, which is less than previously estimated for the ternary complex. Local changes close to the 3-PG binding site include an ordering of Arg-65 and a shift of helix-5.

Original languageEnglish
Pages (from-to)133-144
Number of pages12
JournalProteins: Structure, Function and Genetics
Volume12
Issue number2
Publication statusPublished - 1992

Fingerprint

Phosphoglycerate Kinase
Muscle
Hydrogen
Swine
Crystal structure
Muscles
Substrates
Enzymes
Horses
R388
Phosphates
Binding Sites
Basic Amino Acids
Atoms
Coulomb interactions
Static Electricity
Isomers
Hydroxyl Radical
Hydrogen bonds
Nitrogen

Keywords

  • conformational changes
  • enzyme structure
  • hinge bending
  • substrate binding
  • X-ray crystallography

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Structural Biology

Cite this

Crystal structure of the binary complex of pig muscle phosphoglycerate kinase and its substrate 3-phospho-D-glycerate. / Harlos, K.; Vas, M.; Blake, C. F.

In: Proteins: Structure, Function and Genetics, Vol. 12, No. 2, 1992, p. 133-144.

Research output: Contribution to journalArticle

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