The coiled-coil domain of glycosomal membrane-associated Leishmania donovani PEX14: cloning, overexpression, purification and preliminary crystallographic analysis

The glycosomal membrane-associated Leishmania donovani protein PEX14, which plays a crucial role in protein import from the cytosol to the glycosomal matrix, consists of three domains: an N-terminal domain where the signalling molecule binds, a transmembrane domain and an 84-residue coiled-coil domain (CC) that is responsible for oligomerization. CCs are versatile domains that participate in a variety of functions including supramolecular assembly, cellular signalling and transport. Recombinant PEX14 CC was cloned, overexpressed, affinity-purified with in-column thrombin cleavage and further purified by size-exclusion chromatography. Crystals that diffracted to 1.98   Å resolution were obtained from a condition consisting of 1.4   M sodium citrate tribasic dihydrate, 0.1   M HEPES buffer pH 7.5. The crystals belonged to the monoclinic space group C2, with unit-cell parameters a   =   143.98, b = 32.62, c = 95.62   Å , β = 94.68 ° . Structure determination and characterization are in progress.
Source: Acta Crystallographica Section F - Category: Biochemistry Authors: Tags: Leishmania donovani coiled-coil domain PEX14 peroxisomes glycosomes research communications Source Type: research