Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases

C Reidl; KA Mojorek; J Dang; D Tran; K Jew; M Law; Y Payne; W Minor; Daniel P Becker; ML Kuhn

doi:10.1002/1873-3468

Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases

C Reidl
, KA Mojorek
, J Dang
, D Tran
, K Jew
, M Law
, Y Payne
, W Minor
, Daniel P Becker
, ML Kuhn

Loyola University Chicago
University of Virginia
San Francisco State University

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

Abstract

Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases.

Original language	American English
Journal	FEBS Letters
DOIs	https://doi.org/10.1002/1873-3468
State	Published - Aug 2017

Disciplines

Biochemistry, Biophysics, and Structural Biology
Chemistry

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10.1002/1873-3468License: Unspecified

Cite this

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title = "Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases",

abstract = " Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases.",

author = "C Reidl and KA Mojorek and J Dang and D Tran and K Jew and M Law and Y Payne and W Minor and Becker, \{Daniel P\} and ML Kuhn",

note = "Research Letter Corresponding author Department of Chemistry, Loyola University Chicago, IL, USA Correspondence M. L. Kuhn, Department of Chemistry and Biochemistry, San Francisco State University, 1600 Holloway Ave., San Francisco, CA 94132, USA Fax: +1 415 338 2384 Tel: +1 415 405 2112 E-mail: [email protected] and D. P.",

year = "2017",

month = aug,

doi = "10.1002/1873-3468",

language = "American English",

journal = "FEBS Letters",

}

TY - JOUR

T1 - Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases

AU - Reidl, C

AU - Mojorek, KA

AU - Dang, J

AU - Tran, D

AU - Jew, K

AU - Law, M

AU - Payne, Y

AU - Minor, W

AU - Becker, Daniel P

AU - Kuhn, ML

N1 - Research Letter Corresponding author Department of Chemistry, Loyola University Chicago, IL, USA Correspondence M. L. Kuhn, Department of Chemistry and Biochemistry, San Francisco State University, 1600 Holloway Ave., San Francisco, CA 94132, USA Fax: +1 415 338 2384 Tel: +1 415 405 2112 E-mail: [email protected] and D. P.

PY - 2017/8

Y1 - 2017/8

N2 - Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases.

AB - Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases.

UR - https://doi.org/10.1002/1873-3468.12753

U2 - 10.1002/1873-3468

DO - 10.1002/1873-3468

M3 - Article

JO - FEBS Letters

JF - FEBS Letters

ER -

Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases

Abstract

Disciplines

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