Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, BS-10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, Uttar Pradesh 226031, India.
Acta Crystallogr F Struct Biol Commun. 2020 Oct 1;76(Pt 10):464-468. doi: 10.1107/S2053230X20011127. Epub 2020 Sep 15.
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.
亲环蛋白相关的利什曼原虫蛋白 PEX14 定位于糖基体膜，在细胞质到糖基体基质的蛋白质输入过程中发挥关键作用，由三个结构域组成：一个与信号分子结合的 N 端结构域、一个跨膜结构域和一个负责寡聚化的 84 个残基卷曲螺旋（CC）结构域。CC 是多功能结构域，参与多种功能，包括超分子组装、细胞信号转导和运输。重组 PEX14 CC 经克隆、过表达、在柱上用凝血酶切割亲和纯化，然后通过分子筛层析进一步纯化。从包含 1.4 M 三碱基柠檬酸二水合物和 0.1 M HEPES 缓冲液（pH 值 7.5）的条件下获得了可衍射至 1.98 Å 分辨率的晶体。这些晶体属于单斜晶系，空间群为 C2，晶胞参数分别为 a = 143.98、b = 32.62、c = 95.62 Å，β = 94.68°。结构测定和表征正在进行中。
