Structurally-functional study of mutant proteins – misfolding of cystathionin beta-synthase as a model

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Aleš Hnízda, Viktor Kožich

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1.lékařská fakulta UK a VFN v Praze

Loss of protein stability due to missense mutation is the most common cause for monogenic diseases. Generally, aminoacid substitution in the protein does not often target key functional residues but rather leads to misfolded protein with disrupted native structure.
In our study, we focused on structural mechanisms underlying protein misfolding of cystathionine beta synthase mutant enzymes. We described conformational properties of nine pathogenic variants directly in crude cell extracts by proteolytic approaches using thermolysin. Other nine CBS mutants were successfully purified after expression in E.coli and subsequently characterized by common spectroscopic and biochemical techniques. We found that the altered degree of protein unfolding is a common structural mechanism for the major disease-causing mutation.General methological strategy for assessment of mutations pathogenicity will be discussed.
 
This work was supported by the by the Research Project from the Ministry of Education of the Czech Republic No. MSM0021620806, by the grant No. 7310 from Grant Agency of Charles University in Prague and by the grant SVV262502 from the Charles University in Prague.

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