The unique functional role of the C-H center dot center dot center dot S hydrogen bond in the substrate specificity and enzyme catalysis of type 1 methionine aminopeptidase

Reddi, Ravikumar and Singarapu, Kiran Kumar and Pal, Debnath and Addlagatta, Anthony (2016) The unique functional role of the C-H center dot center dot center dot S hydrogen bond in the substrate specificity and enzyme catalysis of type 1 methionine aminopeptidase. In: MOLECULAR BIOSYSTEMS, 12 (8). pp. 2408-2416.

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Abstract

It is intriguing how nature attains recognition specificity between molecular interfaces where there is no apparent scope for classical hydrogen bonding or polar interactions. Methionine aminopeptidase (MetAP) is one such enzyme where this fascinating conundrum is at play. In this study, we demonstrate that a unique C-H center dot center dot center dot S hydrogen bond exists between the enzyme methionine aminopeptidase (MetAP) and its N-terminal-methionine polypeptide substrate, which allows specific interaction between apparent apolar interfaces, imposing a strict substrate recognition specificity and efficient catalysis, a feature replicated in Type I MetAPs across all kingdoms of life. We evidence this evolutionarily conserved C-H center dot center dot center dot S hydrogen bond through enzyme assays on wild-type and mutant MetAP proteins from Mycobacterium tuberculosis that show a drastic difference in catalytic efficiency. The X-ray crystallographic structure of the methionine bound protein revealed a conserved water bridge and short contacts involving the Met side-chain, a feature also observed in MetAPs from other organisms. Thermal shift assays showed a remarkable 3.3 degrees C increase in melting temperature for methionine bound protein compared to its norleucine homolog, where C-H center dot center dot center dot S interaction is absent. The presence of C-H center dot center dot center dot S hydrogen bonding was also corroborated by nuclear magnetic resonance spectroscopy through a change in chemical shift. Computational chemistry studies revealed the unique role of the electrostatic environment in facilitating the C-H center dot center dot center dot S interaction. The significance of this atypical hydrogen bond is underscored by the fact that the function of MetAP is essential for any living cell.

Item Type:	Journal Article
Publication:	MOLECULAR BIOSYSTEMS
Publisher:	ROYAL SOC CHEMISTRY
Additional Information:	Copy right for this article belongs to the ROYAL SOC CHEMISTRY, THOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLAND
Department/Centre:	Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Date Deposited:	22 Aug 2016 09:48
Last Modified:	22 Aug 2016 09:48
URI:	http://eprints.iisc.ac.in/id/eprint/54502

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