Stabilization of the thermolabile variant S113L of carnitine palmitoyltransferase II.

ABSTRACT:

Objective

Muscle carnitine palmitoyltransferase (CPT) II deficiency, the most common defect of lipid metabolism in muscle, is characterized by attacks of myoglobinuria without persistent muscle weakness.

Methods

His6-N-hCPT2 (wild-type) and His6-N-hCPT2/S113L (variant) were produced recombinantly in prokaryotic host and characterized according to their functional and regulatory properties.

Results

The wild-type and the variant S113L showed the same enzymatic activity and thermostability at 30°C. The mutated enzyme, however, revealed an abnormal thermal destabilization at 40°C and 45°C. This was consistent with an increased flexibility (B-factor) of the variant at 40°C compared with that of the wild-type shown by molecular dynamics analysis. Preincubation of the enzyme

SUBMITTER: Motlagh L 

PROVIDER: S-EPMC4830186 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature