title: Kinetic stability of cystathionine beta-synthase can be modulated by structural analogs of S-adenosylmethionine: Potential approach to pharmacological chaperone therapy for homocystinuria.
abstract: Many pathogenic missense mutations in human cystathionine beta-synthase (CBS) cause misfolding of the mutant enzyme resulting in aggregation or rapid degradation of the protein. Subsequent loss of CBS function leads to CBS-deficient homocystinuria (CBSDH). CBS contains two sets of binding sites for S-adenosylmethionine (SAM) that independently regulate the enzyme activity and kinetically stabilize its regulatory domain. In the present study, we examined the hypothesis that CBS activation may be decoupled from kinetic stabilization and thus CBS regulatory domain can serve as a novel drug target for CBSDH. We determined the effect of SAM and its close structural analogs on CBS activity, their binding to and stabilization of the regulatory domain in the absence and presence of competing SAM. Binding of S-adenosylhomocysteine and sinefungin lead to stabilization of the regulatory domains without activation of CBS. Direct titrations and competition experiments support specific binding of these two SAM analogs to the stabilizing sites. Binding of these two ligands also affects the enzyme proteolysis rate supporting the role of the stabilizing sites in CBS dynamics. Our results indicate that binding of SAM to regulatory and stabilizing sites in CBS may have evolved to display an exquisite thermodynamic and structural specificity towards SAM as well as the ability to transduce the allosteric signal responsible for CBS activation. Thus, ligands may be developed to function as kinetic stabilizers or pharmacological chaperones without interfering with the physiological activation of CBS by SAM.
authors.affiliation: Department of Pediatrics, University of Colorado Denver, Aurora, CO 80045, USA.
authors.forename: Tomas
authors.initials: T
authors.lastname: Majtan
authors.affiliation: Department of Physical Chemistry, Faculty of Sciences, University of Granada, Granada 18071, Spain. Electronic address: angelpey@ugr.es.
authors.forename: Angel L
authors.initials: AL
authors.lastname: Pey
authors.affiliation: Department of Pediatrics, University of Colorado Denver, Aurora, CO 80045, USA. Electronic address: jan.kraus@ucdenver.edu.
authors.forename: Jan P
authors.initials: JP
authors.lastname: Kraus
mesh_terms: D003541:Cystathionine beta-Synthase / Q000378:metabolism*; D004795:Enzyme Stability / Q000187:drug effects; D006712:Homocystinuria / Q000188:drug therapy* / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D012436:S-Adenosylmethionine; D003541:Cystathionine beta-Synthase
keywords: Calorimetry; Cystathionine beta-synthase; Homocystinuria; Kinetic stability; Pharmacological chaperone; S-adenosylmethionine