title: Three-stage assembly of the cysteine synthase complex from Escherichia coli.
abstract: Control of sulfur metabolism in plants and bacteria is linked, in significant measure, to the behavior of the cysteine synthase complex (CSC). The complex is comprised of the two enzymes that catalyze the final steps in cysteine biosynthesis: serine O-acetyltransferase (SAT, EC 2.3.1.30), which produces O-acetyl-L-serine, and O-acetyl-L-serine sulfhydrylase (OASS, EC 2.5.1.47), which converts it to cysteine. SAT (a dimer of homotrimers) binds a maximum of two molecules of OASS (a dimer) in an interaction believed to involve docking of the C terminus from a protomer in an SAT trimer into an OASS active site. This interaction inactivates OASS catalysis and prevents further binding to the trimer; thus, the system exhibits a contact-induced inactivation of half of each biomolecule. To better understand the dynamics and energetics that underlie formation of the CSC, the interactions of OASS and SAT from Escherichia coli were studied at equilibrium and in the pre-steady state. Using an experimental strategy that initiates dissociation of the CSC at different points in the CSC-forming reaction, three stable forms of the complex were identified. Comparison of the binding behaviors of SAT and its C-terminal peptide supports a mechanism in which SAT interacts with OASS in a non-allosteric interaction involving its C terminus. This early docking event appears to fasten the proteins in close proximity and thus prepares the system to engage in a series of subsequent, energetically favorable isomerizations that inactivate OASS and produce the fully isomerized CSC.
authors.affiliation: Department of Microbiology and Immunology, The Albert Einstein College of Medicine, Bronx, New York 10461, USA.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Leyh
mesh_terms: D003545:Cysteine / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D003547:Cysteine Synthase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D048168:Escherichia coli K12 / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020836:Protein Structure, Quaternary
chemical_list: D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D003547:Cysteine Synthase; D003545:Cysteine