Structural snapshots of MTA/AdoHcy nucleosidase along the reaction coordinate provide insights into enzyme and nucleoside flexibility during catalysis

Jeffrey E. Lee; G. David Smith; Cathy Horvatin; David J.T. Huang; Kenneth A. Cornell; Michael K. Riscoe; P. Lynne Howell

doi:10.1016/j.jmb.2005.07.027

Structural snapshots of MTA/AdoHcy nucleosidase along the reaction coordinate provide insights into enzyme and nucleoside flexibility during catalysis

Jeffrey E. Lee
, G. David Smith
, Cathy Horvatin
, David J.T. Huang
, Kenneth A. Cornell
, Michael K. Riscoe
, P. Lynne Howell

Research output: Contribution to journal › Article › peer-review

39 Scopus citations

Abstract

MTA/AdoHcy nucleosidase (MTAN) irreversibly hydrolyzes the N9-C1′ bond in the nucleosides, 5′-methylthioadenosine (MTA) and S-adenosylhomocysteine (AdoHcy) to form adenine and the corresponding thioribose. MTAN plays a vital role in metabolic pathways involving methionine recycling, biological methylation, polyamine biosynthesis, and quorum sensing. Crystal structures of a wild-type (WT) MTAN complexed with glycerol, and mutant-enzyme and mutant-product complexes have been determined at 2.0 Å, 2.0 Å, and 2.1 Å resolution, respectively. The WT MTAN-glycerol structure provides a purine-free model and in combination with the previously solved thioribose-free MTAN-ADE structure, we now have separate apo structures for both MTAN binding subsites. The purine and thioribose-free states reveal an extensive enzyme-immobilized water network in their respective binding subsites. The Asp197Asn MTAN-MTA and Glu12Gln MTAN-MTR·ADE structures are the first enzyme-substrate and enzyme-product complexes reported for MTAN, respectively. These structures provide representative snapshots along the reaction coordinate and allow insight into the conformational changes of the enzyme and the nucleoside substrate. A "catalytic movie" detailing substrate binding, catalysis, and product release is presented.

Original language	English (US)
Pages (from-to)	559-574
Number of pages	16
Journal	Journal of molecular biology
Volume	352
Issue number	3
DOIs	https://doi.org/10.1016/j.jmb.2005.07.027
State	Published - Sep 23 2005
Externally published	Yes

Funding

The authors thank Dr Yuri Lobsanov for his help in data collection at the Hospital for Sick Children X-ray diffraction facility and Quorex Pharmaceuticals (Carlsbad, CA) for providing QX-10000024. This work is supported by grants from the Canadian Institute for Health Research (CIHR, no. 43998), the United States Department of Agriculture (02-0047), the United States Veterans Affairs Medical Research Program, and a sponsored research agreement from Quorex Pharmaceuticals. P.L.H. and D.J.T.H. are supported by a CIHR investigator award and a Natural Sciences and Engineering Research Council of Canada (NSERC) summer studentship, respectively. J.E.L. was supported, fully or in part, by a doctoral research award from CIHR and a studentship from the Ontario Student Opportunities Trust Fund and Hospital for Sick Children Foundation Student Scholarship Program.

Funders	Funder number
IMHA Canadian Institute for Health Research	43998
Ontario Student Opportunities Trust Fund
Quorex Pharmaceuticals
United States Veterans Affairs Medical Research Program
U.S. Department of Agriculture	02-0047
Natural Sciences and Engineering Research Council of Canada
Hospital for Sick Children

Keywords

5′-methylthioadenosine
5′-methylthioadenosine/S- adenosylhomocysteine nucleosidase
Conformational flexibility
Reaction coordinate
S-adenosylhomocysteine

ASJC Scopus subject areas

Molecular Biology
Biophysics
Structural Biology

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10.1016/j.jmb.2005.07.027

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@article{7f1c18c16a34430485b5f1f24fb9c7d0,

title = "Structural snapshots of MTA/AdoHcy nucleosidase along the reaction coordinate provide insights into enzyme and nucleoside flexibility during catalysis",

abstract = "MTA/AdoHcy nucleosidase (MTAN) irreversibly hydrolyzes the N9-C1′ bond in the nucleosides, 5′-methylthioadenosine (MTA) and S-adenosylhomocysteine (AdoHcy) to form adenine and the corresponding thioribose. MTAN plays a vital role in metabolic pathways involving methionine recycling, biological methylation, polyamine biosynthesis, and quorum sensing. Crystal structures of a wild-type (WT) MTAN complexed with glycerol, and mutant-enzyme and mutant-product complexes have been determined at 2.0 {\AA}, 2.0 {\AA}, and 2.1 {\AA} resolution, respectively. The WT MTAN-glycerol structure provides a purine-free model and in combination with the previously solved thioribose-free MTAN-ADE structure, we now have separate apo structures for both MTAN binding subsites. The purine and thioribose-free states reveal an extensive enzyme-immobilized water network in their respective binding subsites. The Asp197Asn MTAN-MTA and Glu12Gln MTAN-MTR·ADE structures are the first enzyme-substrate and enzyme-product complexes reported for MTAN, respectively. These structures provide representative snapshots along the reaction coordinate and allow insight into the conformational changes of the enzyme and the nucleoside substrate. A {"}catalytic movie{"} detailing substrate binding, catalysis, and product release is presented.",

keywords = "5′-methylthioadenosine, 5′-methylthioadenosine/S- adenosylhomocysteine nucleosidase, Conformational flexibility, Reaction coordinate, S-adenosylhomocysteine",

author = "Lee, \{Jeffrey E.\} and Smith, \{G. David\} and Cathy Horvatin and Huang, \{David J.T.\} and Cornell, \{Kenneth A.\} and Riscoe, \{Michael K.\} and Howell, \{P. Lynne\}",

note = "Funding Information: The authors thank Dr Yuri Lobsanov for his help in data collection at the Hospital for Sick Children X-ray diffraction facility and Quorex Pharmaceuticals (Carlsbad, CA) for providing QX-10000024. This work is supported by grants from the Canadian Institute for Health Research (CIHR, no. 43998), the United States Department of Agriculture (02-0047), the United States Veterans Affairs Medical Research Program, and a sponsored research agreement from Quorex Pharmaceuticals. P.L.H. and D.J.T.H. are supported by a CIHR investigator award and a Natural Sciences and Engineering Research Council of Canada (NSERC) summer studentship, respectively. J.E.L. was supported, fully or in part, by a doctoral research award from CIHR and a studentship from the Ontario Student Opportunities Trust Fund and Hospital for Sick Children Foundation Student Scholarship Program. ",

year = "2005",

month = sep,

day = "23",

doi = "10.1016/j.jmb.2005.07.027",

language = "English (US)",

volume = "352",

pages = "559--574",

journal = "Journal of molecular biology",

issn = "0022-2836",

publisher = "Academic Press",

number = "3",

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TY - JOUR

T1 - Structural snapshots of MTA/AdoHcy nucleosidase along the reaction coordinate provide insights into enzyme and nucleoside flexibility during catalysis

AU - Lee, Jeffrey E.

AU - Smith, G. David

AU - Horvatin, Cathy

AU - Huang, David J.T.

AU - Cornell, Kenneth A.

AU - Riscoe, Michael K.

AU - Howell, P. Lynne

N1 - Funding Information: The authors thank Dr Yuri Lobsanov for his help in data collection at the Hospital for Sick Children X-ray diffraction facility and Quorex Pharmaceuticals (Carlsbad, CA) for providing QX-10000024. This work is supported by grants from the Canadian Institute for Health Research (CIHR, no. 43998), the United States Department of Agriculture (02-0047), the United States Veterans Affairs Medical Research Program, and a sponsored research agreement from Quorex Pharmaceuticals. P.L.H. and D.J.T.H. are supported by a CIHR investigator award and a Natural Sciences and Engineering Research Council of Canada (NSERC) summer studentship, respectively. J.E.L. was supported, fully or in part, by a doctoral research award from CIHR and a studentship from the Ontario Student Opportunities Trust Fund and Hospital for Sick Children Foundation Student Scholarship Program.

PY - 2005/9/23

Y1 - 2005/9/23

N2 - MTA/AdoHcy nucleosidase (MTAN) irreversibly hydrolyzes the N9-C1′ bond in the nucleosides, 5′-methylthioadenosine (MTA) and S-adenosylhomocysteine (AdoHcy) to form adenine and the corresponding thioribose. MTAN plays a vital role in metabolic pathways involving methionine recycling, biological methylation, polyamine biosynthesis, and quorum sensing. Crystal structures of a wild-type (WT) MTAN complexed with glycerol, and mutant-enzyme and mutant-product complexes have been determined at 2.0 Å, 2.0 Å, and 2.1 Å resolution, respectively. The WT MTAN-glycerol structure provides a purine-free model and in combination with the previously solved thioribose-free MTAN-ADE structure, we now have separate apo structures for both MTAN binding subsites. The purine and thioribose-free states reveal an extensive enzyme-immobilized water network in their respective binding subsites. The Asp197Asn MTAN-MTA and Glu12Gln MTAN-MTR·ADE structures are the first enzyme-substrate and enzyme-product complexes reported for MTAN, respectively. These structures provide representative snapshots along the reaction coordinate and allow insight into the conformational changes of the enzyme and the nucleoside substrate. A "catalytic movie" detailing substrate binding, catalysis, and product release is presented.

AB - MTA/AdoHcy nucleosidase (MTAN) irreversibly hydrolyzes the N9-C1′ bond in the nucleosides, 5′-methylthioadenosine (MTA) and S-adenosylhomocysteine (AdoHcy) to form adenine and the corresponding thioribose. MTAN plays a vital role in metabolic pathways involving methionine recycling, biological methylation, polyamine biosynthesis, and quorum sensing. Crystal structures of a wild-type (WT) MTAN complexed with glycerol, and mutant-enzyme and mutant-product complexes have been determined at 2.0 Å, 2.0 Å, and 2.1 Å resolution, respectively. The WT MTAN-glycerol structure provides a purine-free model and in combination with the previously solved thioribose-free MTAN-ADE structure, we now have separate apo structures for both MTAN binding subsites. The purine and thioribose-free states reveal an extensive enzyme-immobilized water network in their respective binding subsites. The Asp197Asn MTAN-MTA and Glu12Gln MTAN-MTR·ADE structures are the first enzyme-substrate and enzyme-product complexes reported for MTAN, respectively. These structures provide representative snapshots along the reaction coordinate and allow insight into the conformational changes of the enzyme and the nucleoside substrate. A "catalytic movie" detailing substrate binding, catalysis, and product release is presented.

KW - 5′-methylthioadenosine

KW - 5′-methylthioadenosine/S- adenosylhomocysteine nucleosidase

KW - Conformational flexibility

KW - Reaction coordinate

KW - S-adenosylhomocysteine

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UR - https://www.scopus.com/pages/publications/24044453721#tab=citedBy

U2 - 10.1016/j.jmb.2005.07.027

DO - 10.1016/j.jmb.2005.07.027

M3 - Article

C2 - 16109423

AN - SCOPUS:24044453721

SN - 0022-2836

VL - 352

SP - 559

EP - 574

JO - Journal of molecular biology

JF - Journal of molecular biology

IS - 3

ER -

Structural snapshots of MTA/AdoHcy nucleosidase along the reaction coordinate provide insights into enzyme and nucleoside flexibility during catalysis

Abstract

Funding

Keywords

ASJC Scopus subject areas

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