@inbook{85c46bf797994eab895cbf24959e3a77,
title = "Structural characterization of cobalamin-dependent radical S-adenosylmethionine methylases",
abstract = "Cobalamin-dependent radical S-adenosylmethionine (SAM) methylases catalyze key steps in the biosynthesis of numerous biomolecules, including protein cofactors, antibiotics, herbicides, and other natural products, but have remained a relatively understudied subclass of radical SAM enzymes due to their inherent insolubility upon overproduction in Escherichia coli. These enzymes contain two cofactors: a [4Fe-4S] cluster that is ligated by three cysteine residues, and a cobalamin cofactor typically bound by residues in the N-terminal portion of the enzyme. Recent advances in the expression and purification of these enzymes in their active states and with both cofactors present has allowed for more detailed biochemical studies as well as structure determination by X-ray crystallography. Herein, we use KsTsrM and TokK to highlight methods for the structural characterization of cobalamin-dependent radical SAM (RS) enzymes and describe recent advances in in the overproduction and purification of these enzymes.",
author = "Knox, {Hayley L.} and Booker, {Squire J.}",
note = "Funding Information: This work was supported by NIH (GM122595 to S.J.B.) and the Eberly Family Distinguished Chair in Science (S.J.B.). S.J.B. is an investigator of the Howard Hughes Medical Institute. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract no. DE-AC02-06CH11357. Use of GM/CA@APS has been funded in whole or in part with federal funds from the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006). The Eiger 16 M detector at GM/CA-XSD was funded by NIH grant S10 OD012289. Use of the LS-CAT Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (grant 085P1000817). This research also used the resources of the Berkeley Center for Structural Biology supported in part by the Howard Hughes Medical Institute. The Advanced Light Source is a DOE Office of Science User Facility under contract no. DE-AC02-05CH11231. The ALS-ENABLE beamlines are supported in part by the National Institutes of Health, National Institute of General Medical Sciences grant P30 GM124169. Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",
year = "2022",
month = jan,
doi = "10.1016/bs.mie.2021.12.013",
language = "English (US)",
isbn = "9780323955577",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "3--27",
editor = "Marsh, {E. Neil G.}",
booktitle = "Coenzyme B Enzymes Part B",
address = "United States",
}