Active-Site Controlled, Jahn-Teller Enabled Regioselectivity in Reductive S-C Bond Cleavage of S-Adenosylmethionine in Radical SAM Enzymes

J Am Chem Soc. 2021 Jan 13;143(1):335-348. doi: 10.1021/jacs.0c10925. Epub 2020 Dec 29.

Abstract

Catalysis by canonical radical S-adenosyl-l-methionine (SAM) enzymes involves electron transfer (ET) from [4Fe-4S]+ to SAM, generating an R3S0 radical that undergoes regioselective homolytic reductive cleavage of the S-C5' bond to generate the 5'-dAdo· radical. However, cryogenic photoinduced S-C bond cleavage has regioselectively yielded either 5'-dAdo· or ·CH3, and indeed, each of the three SAM S-C bonds can be regioselectively cleaved in an RS enzyme. This diversity highlights a longstanding central question: what controls regioselective homolytic S-C bond cleavage upon SAM reduction? We here provide an unexpected answer, founded on our observation that photoinduced S-C bond cleavage in multiple canonical RS enzymes reveals two enzyme classes: in one, photolysis forms 5'-dAdo·, and in another it forms ·CH3. The identity of the cleaved S-C bond correlates with SAM ribose conformation but not with positioning and orientation of the sulfonium center relative to the [4Fe-4S] cluster. We have recognized the reduced-SAM R3S0 radical is a (2E) state with its antibonding unpaired electron in an orbital doublet, which renders R3S0 Jahn-Teller (JT)-active and therefore subject to vibronically induced distortion. Active-site forces induce a JT distortion that localizes the odd electron in a single priority S-C antibond, which undergoes regioselective cleavage. In photolytic cleavage those forces act through control of the ribose conformation and are transmitted to the sulfur via the S-C5' bond, but during catalysis thermally induced conformational changes that enable ET from a cluster iron generate dominant additional forces that specifically select S-C5' for cleavage. This motion also can explain how 5'-dAdo· subsequently forms the organometallic intermediate Ω.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / radiation effects
  • Biocatalysis
  • Catalytic Domain
  • Clostridium acetobutylicum / enzymology
  • Density Functional Theory
  • Iron-Sulfur Proteins / chemistry
  • Iron-Sulfur Proteins / radiation effects
  • Light
  • Models, Chemical
  • Molecular Structure
  • Oxidation-Reduction / radiation effects
  • Oxidoreductases Acting on Sulfur Group Donors / chemistry*
  • Oxidoreductases Acting on Sulfur Group Donors / radiation effects
  • Photolysis
  • S-Adenosylmethionine / chemistry*
  • S-Adenosylmethionine / radiation effects
  • Thermotoga maritima / enzymology

Substances

  • Bacterial Proteins
  • Iron-Sulfur Proteins
  • S-Adenosylmethionine
  • Oxidoreductases Acting on Sulfur Group Donors