Critical hydrogen bonds and protonation states of pyridoxal 5′-phosphate revealed by NMR

Hans Heinrich Limbach, Monique Chan-Huot, Shasad Sharif, Peter M. Tolstoy, Ilya G. Shenderovich, Gleb S. Denisov, Michael D. Toney

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

In this contribution we review recent NMR studies of protonation and hydrogen bond states of pyridoxal 5′-phosphate (PLP) and PLP model Schiff bases in different environments, starting from aqueous solution, the organic solid state to polar organic solution and finally to enzyme environments. We have established hydrogen bond correlations that allow one to estimate hydrogen bond geometries from 15N chemical shifts. It is shown that protonation of the pyridine ring of PLP in aspartate aminotransferase (AspAT) is achieved by (i) an intermolecular OHN hydrogen bond with an aspartate residue, assisted by the imidazole group of a histidine side chain and (ii) a local polarity as found for related model systems in a polar organic solvent exhibiting a dielectric constant of about 30. Model studies indicate that protonation of the pyridine ring of PLP leads to a dominance of the ketoenamine form, where the intramolecular OHN hydrogen bond of PLP exhibits a zwitterionic state. Thus, the PLP moiety in AspAT carries a net positive charge considered as a pre-requisite to initiate the enzyme reaction. However, it is shown that the ketoenamine form dominates in the absence of ring protonation when PLP is solvated by polar groups such as water. Finally, the differences between acid-base interactions in aqueous solution and in the interior of proteins are discussed. This article is part of a special issue entitled: Pyridoxal Phosphate Enzymology.

Original languageEnglish (US)
Pages (from-to)1425-1437
Number of pages13
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1814
Issue number11
DOIs
StatePublished - Nov 2011

Fingerprint

Pyridoxal Phosphate
Protonation
Hydrogen
Hydrogen bonds
Nuclear magnetic resonance
Aspartate Aminotransferases
Schiff Bases
Chemical shift
Enzymes
Histidine
Aspartic Acid
Organic solvents
Permittivity
Acids
Geometry
Water

Keywords

  • Aspartate aminotransferase
  • Hydrogen bonding
  • Protonation state
  • Pyridoxal 5′-phosphate
  • Solid and liquid state NMR
  • Tautomerism

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Analytical Chemistry
  • Molecular Biology
  • Medicine(all)

Cite this

Limbach, H. H., Chan-Huot, M., Sharif, S., Tolstoy, P. M., Shenderovich, I. G., Denisov, G. S., & Toney, M. D. (2011). Critical hydrogen bonds and protonation states of pyridoxal 5′-phosphate revealed by NMR. Biochimica et Biophysica Acta - Proteins and Proteomics, 1814(11), 1425-1437. https://doi.org/10.1016/j.bbapap.2011.06.004

Critical hydrogen bonds and protonation states of pyridoxal 5′-phosphate revealed by NMR. / Limbach, Hans Heinrich; Chan-Huot, Monique; Sharif, Shasad; Tolstoy, Peter M.; Shenderovich, Ilya G.; Denisov, Gleb S.; Toney, Michael D.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1814, No. 11, 11.2011, p. 1425-1437.

Research output: Contribution to journalArticle

Limbach, HH, Chan-Huot, M, Sharif, S, Tolstoy, PM, Shenderovich, IG, Denisov, GS & Toney, MD 2011, 'Critical hydrogen bonds and protonation states of pyridoxal 5′-phosphate revealed by NMR', Biochimica et Biophysica Acta - Proteins and Proteomics, vol. 1814, no. 11, pp. 1425-1437. https://doi.org/10.1016/j.bbapap.2011.06.004
Limbach, Hans Heinrich ; Chan-Huot, Monique ; Sharif, Shasad ; Tolstoy, Peter M. ; Shenderovich, Ilya G. ; Denisov, Gleb S. ; Toney, Michael D. / Critical hydrogen bonds and protonation states of pyridoxal 5′-phosphate revealed by NMR. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2011 ; Vol. 1814, No. 11. pp. 1425-1437.
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