Evidence for pentagalloyl glucose binding to human salivary α-amylase through aromatic amino acid residues

G. Gyémánt, Ágnes Zajácz, Bálint Bécsi, Chandran Ragunath, Narayanan Ramasubbu, F. Erdődi, G. Batta, L. Kandra

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

We demonstrate here that pentagalloyl glucose (PGG), a main component of gallotannins, was an effective inhibitor of HSA and it exerted similar inhibitory potency to Aleppo tannin used in this study. The inhibition of HSA by PGG was found to be non-competitive and inhibitory constants of KEI = 2.6 μM and KESI = 3.9 μM were determined from Lineweaver-Burk secondary plots. PGG as a model compound for gallotannins was selected to study the inhibitory mechanism and to characterize the interaction of HSA with this type of molecules. Surface plasmon resonance (SPR) binding experiments confirmed the direct interaction of HSA and PGG, and it also established similar binding of Aleppo tannin to HSA. Saturation transfer difference (STD) experiment by NMR clearly demonstrated the aromatic rings of PGG may be involved in the interaction suggesting a possible stacking with the aromatic side chains of HSA. The role of aromatic amino acids of HSA in PGG binding was reinforced by kinetic studies with the W58L and Y151M mutants of HSA: the replacement of the active site aromatic amino acids with aliphatic ones decreased the PGG inhibition dramatically, which justified the importance of these residues in the interaction.

Original languageEnglish
Pages (from-to)291-296
Number of pages6
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1794
Issue number2
DOIs
Publication statusPublished - Feb 2009

Fingerprint

Aromatic Amino Acids
Amylases
Glucose
Hydrolyzable Tannins
Surface Plasmon Resonance
Surface plasmon resonance
Catalytic Domain
Experiments
Nuclear magnetic resonance
Molecules
Kinetics

Keywords

  • Human salivary α-amylase inhibition
  • NMR
  • Pentagalloyl glucose
  • Saturation transfer difference
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Analytical Chemistry
  • Molecular Biology

Cite this

Evidence for pentagalloyl glucose binding to human salivary α-amylase through aromatic amino acid residues. / Gyémánt, G.; Zajácz, Ágnes; Bécsi, Bálint; Ragunath, Chandran; Ramasubbu, Narayanan; Erdődi, F.; Batta, G.; Kandra, L.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1794, No. 2, 02.2009, p. 291-296.

Research output: Contribution to journalArticle

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AU - Ramasubbu, Narayanan

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