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The coiled‐coil domain of glycosomal membrane‐associated Leishmania donovani PEX14: cloning, overexpression, purification and preliminary crystallographic analysis
Acta Crystallographica Section F ( IF 1.1 ) Pub Date : 2020-10-02 , DOI: 10.1107/s2053230x20011127 Anil Kumar Shakya 1 , J Venkatesh Pratap 1
Acta Crystallographica Section F ( IF 1.1 ) Pub Date : 2020-10-02 , DOI: 10.1107/s2053230x20011127 Anil Kumar Shakya 1 , J Venkatesh Pratap 1
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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.
中文翻译：
糖体膜相关利什曼原虫 PEX14 的卷曲螺旋结构域：克隆、过表达、纯化和初步晶体学分析
糖体膜相关利什曼原虫蛋白 PEX14 在蛋白质从细胞质到糖体基质的输入中起关键作用，由三个结构域组成：信号分子结合的 N 端结构域、跨膜结构域和 84 个残基卷曲螺旋结构域 (CC) 负责寡聚化。CCs 是参与多种功能的多功能域，包括超分子组装、细胞信号传导和运输。重组 PEX14 CC 被克隆、过表达、用柱内凝血酶裂解亲和纯化，并通过尺寸排阻色谱进一步纯化。衍射至 1.98 Å 分辨率的晶体是从由 1.4 M柠檬酸三钠二水合物、0.1 MHEPES 缓冲液 pH 7.5。晶体属于单斜空间群C 2，晶胞参数a = 143.98, b = 32.62, c = 95.62 Å, β = 94.68°。结构确定和表征正在进行中。
更新日期：2020-10-02
中文翻译：
糖体膜相关利什曼原虫 PEX14 的卷曲螺旋结构域：克隆、过表达、纯化和初步晶体学分析
糖体膜相关利什曼原虫蛋白 PEX14 在蛋白质从细胞质到糖体基质的输入中起关键作用，由三个结构域组成：信号分子结合的 N 端结构域、跨膜结构域和 84 个残基卷曲螺旋结构域 (CC) 负责寡聚化。CCs 是参与多种功能的多功能域，包括超分子组装、细胞信号传导和运输。重组 PEX14 CC 被克隆、过表达、用柱内凝血酶裂解亲和纯化，并通过尺寸排阻色谱进一步纯化。衍射至 1.98 Å 分辨率的晶体是从由 1.4 M柠檬酸三钠二水合物、0.1 MHEPES 缓冲液 pH 7.5。晶体属于单斜空间群C 2，晶胞参数a = 143.98, b = 32.62, c = 95.62 Å, β = 94.68°。结构确定和表征正在进行中。
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