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Restoring assembly and activity of cystathionine β-synthase mutants by ligands and chemical chaperones
J. Kopecká, J. Krijt, K. Raková, V. Kožich
Language English Country Netherlands
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
NLK
ProQuest Central
from 1999-02-01 to 2018-11-30
Medline Complete (EBSCOhost)
from 2009-08-01 to 1 year ago
Health & Medicine (ProQuest)
from 1999-02-01 to 2018-11-30
- MeSH
- Alleles MeSH
- Betaine pharmacology therapeutic use MeSH
- Cystathionine beta-Synthase chemistry drug effects genetics metabolism MeSH
- Escherichia coli metabolism MeSH
- Glycerol pharmacology MeSH
- Homocystinuria drug therapy genetics metabolism MeSH
- Polymorphism, Single Nucleotide physiology MeSH
- Protein Conformation drug effects MeSH
- Aminolevulinic Acid pharmacology therapeutic use MeSH
- Humans MeSH
- Ligands MeSH
- Molecular Chaperones pharmacology therapeutic use MeSH
- Protein Multimerization drug effects MeSH
- Mutant Proteins chemistry drug effects metabolism MeSH
- Protein Folding drug effects MeSH
- Taurine pharmacology therapeutic use MeSH
- Protein Binding MeSH
- Structure-Activity Relationship MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Misfolding and aggregation of mutant enzymes have been proposed to play role in the pathogenesis of homocystinuria due to cystathionine β-synthase (CBS) deficiency. Chemical chaperones have been recently shown to facilitate proper assembly of several CBS mutants. To asses the number of patients that may respond to chaperone therapy, we examined the effect of selected CBS ligands and osmolytes on assembly and activity of 27 CBS mutants that represent 70% of known CBS alleles. The mutant enzymes were expressed in a bacterial system, and their properties were assessed by native Western blotting and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) assay, respectively. We studied the chaperoning activity of δ-aminolevulinic acid (δ-ALA)-a heme precursor-and of three osmolytes betaine, 2-aminoethanesulfonic acid (taurine), and glycerol. Fourteen mutants responded by at least 30% increase in the amount of correctly assembled tetramers and enzymatic activity to the coexpressional presence of either 0.5 mM δ-ALA, 100 mM betaine, and/or 750 mM glycerol. Eight of these mutants (p.R266K, p.P49L, p.R125Q, p.K102N, p.R369C, p.V180A, p.P78R, p.S466L) were rescuable by all of these three substances. Four mutants showed increased formation of tetramers that was not accompanied by changes in activity. Topology of mutations appeared to determine the chaperone responsiveness, as 11 of 14 solvent-exposed mutations were substantially more responsive than three of 13 buried mutations. This study identified chaperone-responsive mutants that represent 56 of 713 known patient-derived CBS alleles and may serve as a basis for exploring pharmacological approaches aimed at correcting misfolding in homocystinuria.
References provided by Crossref.org
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