Structural basis for the substrate selectivity of a HAD phosphatase from Thermococcus onnurineus NA1 SCIE SCOPUS

Cited 1 time in WEB OF SCIENCE Cited 1 time in Scopus
Title
Structural basis for the substrate selectivity of a HAD phosphatase from Thermococcus onnurineus NA1
Author(s)
Tri Duc Ngo; Binh Van Le; Subramani, Vinod Kumar; Chi My Thi Nguyen; Lee, Hyun Sook; Cho, Yona; Kim, Kyeong Kyu; Hwang, Hye-Yeon
KIOST Author(s)
Lee, Hyun Sook(이현숙)
Publication Year
2015-05-22
Abstract
Proteins in the haloalkaloic acid dehalogenase (HAD) superfamily, which is one of the largest enzyme families, is generally composed of a catalytic core domain and a cap domain. Although proteins in this family show broad substrate specificities, the mechanisms of their substrate recognition are not well understood. In this study, we identified a new substrate binding motif of HAD proteins from structural and functional analyses, and propose that this motif might be crucial for interacting with hydrophobic rings of substrates. The crystal structure of TON_0338, one of the 17 putative HAD proteins identified in a hyperthermophilic archaeon, Thermo coccus onnurineus NA1, was determined as an apo-form at 2.0 A resolution. In addition, we determined the crystal structure TON_0338 in complex with Mg2+ or N-cyclohexyl-2-aminoethanesulfonic acid (CHES) at 1.7 angstrom resolution. Examination of the apo-form and CHES-bound structures revealed that CHES is sandwiched between Trp58 and Trp61, suggesting that this Trp sandwich might function as a substrate recognition motif. In the phosphatase assay, TON_0338 was shown to have high activity for flavin mononucleotide (FMN), and the docking analysis suggested that the flavin of FMN may interact with Trp58 and Trp61 in a way similar to that observed in the crystal structure. Moreover, the replacement of these tryptophan residues significantly reduced the phosphatase activity for FMN. Our results suggest that WxxW may function as a substrate binding motif in HAD proteins, and expand the diversity of their substrate recognition mode. (C) 2015 Elsevier Inc. All rights reserved.
ISSN
0006-291X
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/2486
DOI
10.1016/j.bbrc.2015.03.179
Bibliographic Citation
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, v.461, no.1, pp.122 - 127, 2015
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Subject
BETA-PHOSPHOGLUCOMUTASE; SUPERFAMILY; DEHALOGENASE; SPECIFICITY
Keywords
HAD; Phosphatase; Substrate recognition motif; Crystal structure
Type
Article
Language
English
Document Type
Article
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Related Researcher
Research Interests

Microbial biotechnology,Marine enzymes,미생물 활용,효소 개발

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