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Published October 13, 2009 | Accepted Version + Supplemental Material
Journal Article Open

Arginine, a Key Residue for the Enhancing Ability of an Antifreeze Protein of the Beetle Dendroides canadensis

Abstract

Antifreeze proteins (AFPs) can produce a difference between the nonequilibrium freezing point and the melting point, termed thermal hysteresis (TH). The TH activity of an antifreeze protein (AFP) depends on the specific AFP and its concentration as well as the presence of cosolutes including low molecular mass solutes and/or proteins. We recently identified series of carboxylates and polyols as efficient enhancers for an AFP from the beetle Dendroides canadensis. In this study, we chemically modified DAFP-1 using the arginine-specific reagent 1,2-cyclohexanedione. We demonstrated that 1,2-cyclohexanedione specifically modifies one arginine residue and the modified DAFP-1 loses its enhancing ability completely or partially in the presence of previously identified enhancers. The stronger the enhancement ability of the enhancer on the native DAFP-1, the stronger the enhancement effect of the enhancer on the modified DAFP-1. The weaker enhancers (e.g., glycerol) completely lose their enhancement effect on the modified DAFP-1 due to their inability to compete with 1,2-cyclohexanedione for the arginine residue. Regeneration of the arginine residue using hydroxylamine fully restored the enhancing ability of DAFP-1. These studies indicated that an arginine residue is critical for the enhancing ability of DAFP-1 and the guanidinium group of the arginine residue is important for its interaction with the enhancers, where the general mechanism of arginine−ligand interaction is borne. This work may initiate a complete mechanistic study of the enhancement effect in AFPs.

Additional Information

© 2009 American Chemical Society. Received July 26, 2009; Revised Manuscript Received September 6, 2009. Publication Date (Web): September 11, 2009. This study was supported by National Institutes of Health Grant 1SC3GM086249-01 and the NIH-RIMI program at California State University, Los Angeles (P20 MD001824-01). We thank the Protein and Nucleic Acid Facility at Stanford University, Stanford, CA, for MALDI-TOF mass spectrometry. VMD was developed by the Theoretical and Computational Biophysics Group in the Beckman Institute for Advanced Science and Technology at the University of Illinois at Urbana−Champaign. Supporting Information: HPLC profiles of the modification reaction of DAFP-1, the antifreeze activities of DAFP-1and the Arg-modified DAFP-1 in the presence of various enhancers, and MALDI-TOF mass spectrum of DAFP-1 incubated with citrate. This material is available free of charge via the Internet at http://pubs.acs.org.

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Accepted Version - nihms-147193.pdf

Supplemental Material - bi901283p_si_001.pdf

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Additional details

Created:
August 19, 2023
Modified:
October 19, 2023