Evolutionary repurposing of a sulfatase: A new Michaelis complex leads to efficient transition state charge offset

Proceedings of the National Academy of Sciences of the United States of America
Charlotte M MitonFlorian Hollfelder

Abstract

The recruitment and evolutionary optimization of promiscuous enzymes is key to the rapid adaptation of organisms to changing environments. Our understanding of the precise mechanisms underlying enzyme repurposing is, however, limited: What are the active-site features that enable the molecular recognition of multiple substrates with contrasting catalytic requirements? To gain insights into the molecular determinants of adaptation in promiscuous enzymes, we performed the laboratory evolution of an arylsulfatase to improve its initially weak phenylphosphonate hydrolase activity. The evolutionary trajectory led to a 100,000-fold enhancement of phenylphosphonate hydrolysis, while the native sulfate and promiscuous phosphate mono- and diester hydrolyses were only marginally affected (≤50-fold). Structural, kinetic, and in silico characterizations of the evolutionary intermediates revealed that two key mutations, T50A and M72V, locally reshaped the active site, improving access to the catalytic machinery for the phosphonate. Measured transition state (TS) charge changes along the trajectory suggest the creation of a new Michaelis complex (E•S, enzyme-substrate), with enhanced leaving group stabilization in the TS for the promiscuous ...Continue Reading

References

Jan 1, 1976·Annual Review of Microbiology·R A Jensen
Jun 10, 1977·Science·F Jacob
Nov 19, 1974·Proceedings of the Royal Society of London. Series B, Containing Papers of a Biological Character·A R Fersht
Jul 1, 1997·Protein Science : a Publication of the Protein Society·S D Rader, D A Agard
Apr 1, 1999·Chemistry & Biology·P J O'Brien, D Herschlag
May 15, 2002·The EMBO Journal·Desiree H Fong, Albert M Berghuis
Jan 2, 2003·Proceedings of the National Academy of Sciences of the United States of America·Gerwald JoglLiang Tong
Apr 26, 2003·Current Opinion in Chemical Biology·Shelley D Copley
Jul 25, 2003·Trends in Biochemical Sciences·Leo C James, Dan S Tawfik
Sep 24, 2004·Journal of the American Chemical Society·Ivana Nikolic-HughesDaniel Herschlag
Oct 20, 2004·Angewandte Chemie·Sarah R HansonChi-Huey Wong
Nov 30, 2004·Nature Genetics·Amir AharoniDan S Tawfik
Apr 6, 2005·Biochemistry·Stephen McQ Gould, Dan S Tawfik
Jan 26, 2006·Journal of the American Chemical Society·Jesse G Zalatan, Daniel Herschlag
Apr 19, 2006·Immunity·Roy A Mariuzza
Aug 10, 2006·Chemical Reviews·W Wallace Cleland, Alvan C Hengge
Sep 7, 2006·Proceedings of the National Academy of Sciences of the United States of America·Marika Ekroos, Tove Sjögren
Mar 24, 2007·Trends in Biotechnology·Karl Hult, Per Berglund
Apr 24, 2007·Angewandte Chemie·Miguel D ToscanoDonald Hilvert
Dec 14, 2007·Nature·Katherine Henzler-Wildman, Dorothee Kern
Dec 18, 2007·Journal of Molecular Biology·Colin J JacksonDavid L Ollis
Feb 27, 2008·Biochemistry·Stephen J BenkovicSharon Hammes-Schiffer
May 23, 2008·Journal of Molecular Biology·Shimon BershteinDan S Tawfik
Jul 16, 2008·Journal of Molecular Biology·Rudi FasanFrances H Arnold
Sep 2, 2008·Proceedings of the National Academy of Sciences of the United States of America·Sean Yu McLoughlin, Shelley D Copley
Sep 19, 2008·Journal of the American Chemical Society·Claire McWhirterNicholas H Williams
Feb 11, 2009·Nature Biotechnology·Irene NobeliJanet M Thornton
Apr 11, 2009·Science·Nobuhiko Tokuriki, Dan S Tawfik
Apr 18, 2009·Angewandte Chemie·Ann C BabtieFlorian Hollfelder
Jul 22, 2009·Nature Chemical Biology·Shelley D Copley
Oct 21, 2009·Nature Chemical Biology·David D BoehrPeter E Wright
Jan 19, 2010·Current Opinion in Chemical Biology·Ann BabtieFlorian Hollfelder
Mar 20, 2010·Annual Review of Biochemistry·Olga Khersonsky, Dan S Tawfik
Sep 9, 2010·Current Opinion in Chemical Biology·Buyong Ma, Ruth Nussinov

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Citations

Apr 7, 2020·The FEBS Journal·Shelley D Copley
Dec 12, 2019·Natural Product Reports·Marc G ChevretteFrancisco Barona-Gómez
Oct 23, 2019·Nature Chemical Biology·Gloria YangNobuhiko Tokuriki
Jun 20, 2020·Protein Science : a Publication of the Protein Society·Gloria YangNobuhiko Tokuriki
Dec 7, 2018·International Journal of Molecular Sciences·Tushar Modi, S Banu Ozkan
Jan 30, 2021·Biotechnology Advances·Joan Planas-IglesiasDavid Bednar
Jan 6, 2019·Current Opinion in Biotechnology·Devin L Trudeau, Dan S Tawfik
Jun 5, 2020·Journal of the American Chemical Society·Rory M CreanShina C L Kamerlin
Jun 3, 2021·Current Opinion in Structural Biology·Charlotte M MitonNobuhiko Tokuriki

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