Why Nature Uses Radical SAM Enzymes so Widely: Electron Nuclear Double Resonance Studies of Lysine 2,3-Aminomutase Show the 5'-dAdo⢠"Free Radical" Is Never Free.
J Am Chem Soc
; 137(22): 7111-21, 2015 Jun 10.
Article
em En
| MEDLINE
| ID: mdl-25923449
ABSTRACT
Lysine 2,3-aminomutase (LAM) is a radical S-adenosyl-L-methionine (SAM) enzyme and, like other members of this superfamily, LAM utilizes radical-generating machinery comprising SAM anchored to the unique Fe of a [4Fe-4S] cluster via a classical five-membered N,O chelate ring. Catalysis is initiated by reductive cleavage of the SAM S-C5' bond, which creates the highly reactive 5'-deoxyadenosyl radical (5'-dAdoâ¢), the same radical generated by homolytic Co-C bond cleavage in B12 radical enzymes. The SAM surrogate S-3',4'-anhydroadenosyl-L-methionine (anSAM) can replace SAM as a cofactor in the isomerization of L-α-lysine to L-ß-lysine by LAM, via the stable allylic anhydroadenosyl radical (anAdoâ¢). Here electron nuclear double resonance (ENDOR) spectroscopy of the anAdo⢠radical in the presence of (13)C, (2)H, and (15)N-labeled lysine completes the picture of how the active site of LAM from Clostridium subterminale SB4 "tames" the 5'-dAdo⢠radical, preventing it from carrying out harmful side reactions this "free radical" in LAM is never free. The low steric demands of the radical-generating [4Fe-4S]/SAM construct allow the substrate target to bind adjacent to the S-C5' bond, thereby enabling the 5'-dAdo⢠radical created by cleavage of this bond to react with its partners by undergoing small motions, â¼0.6 Å toward the target and â¼1.5 Å overall, that are controlled by tight van der Waals contact with its partners. We suggest that the accessibility to substrate and ready control of the reactive C5' radical, with "van der Waals control" of small motions throughout the catalytic cycle, is common within the radical SAM enzyme superfamily and is a major reason why these enzymes are the preferred means of initiating radical reactions in nature.
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Base de dados:
MEDLINE
Assunto principal:
Espectroscopia de Ressonância de Spin Eletrônica
/
Transferases Intramoleculares
/
Hidrolases
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article