Structural Exploration on Palmitoyltransferase DHHC3 from Homo sapiens.
Structural Exploration on Palmitoyltransferase DHHC3 from Homo sapiens.
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智人棕榈酰转移酶 DHHC3 的结构探索
作者：
DHHC3属于DHHC棕榈酰转移酶家族，该家族通过在半胱氨酸上连接一个脂肪酰基来催化靶蛋白的s -棕榈酰化。近年来，DHHC3已被证明是一种有前景的抗肿瘤靶点。然而，考虑到其序列与已知晶体结构的DHHC同源物的差异，DHHC3的详细结构和催化机制尚不清楚。在这里，我们描述了人DHHC3 （hDHHC3）的表达和纯化，并截断了hDHHC3的柔性n端结构域（NTD）。纯化的hDHHC3蛋白在不同条件下进行蛋白结晶。LAMTOR1是与hDHHC3相互作用促进结晶的蛋白之一，通过质谱和共免疫沉淀法进一步鉴定。利用低温电镜对失活hDHHS3突变体进行结构探索，显示出典型的膜蛋白侧视图。这些结果为DHHC3的结构测定提供了初步的指导。
DHHC3 belongs to a family of DHHC palmitoyltransferase, which catalyzes the S-palmitoylation of target proteins by attaching a fatty acyl group to a cysteine. Recently, DHHC3 has been demonstrated to be a promising antitumor target in cancer therapeutics. However, the detailed structure and catalysis mechanism of DHHC3 remain elusive, considering its sequence diversity from the DHHC homologues with known crystal structures. Here, we described the expression and purification of human DHHC3 (hDHHC3) and truncated hDHHC3 with the flexible N-terminal domain (NTD) removed. Purified hDHHC3 proteins were used under various conditions for protein crystallization. LAMTOR1, one of the interacting proteins of hDHHC3 to facilitate the crystallization, was further identified by mass spectrometry and co-immunoprecipitation assay. The structural exploration using cryogenic electronic microscopy (cryo-EM) on the inactive hDHHS3 mutant showed a typical sideview of membrane proteins. These results provide a preliminary guidance for the structural determination of DHHC3.
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DOI：
10.1073/pnas.2022050119
发表时间：
2022-02-15
影响因子：
11.1
作者：
Lee CJ;Stix R;Rana MS;Shikwana F;Murphy RE;Ghirlando R;Faraldo-Gómez JD;Banerjee A
通讯作者：
Banerjee A
DOI：
10.1126/science.aao6326
发表时间：
2018-01-12
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Rana MS;Kumar P;Lee CJ;Verardi R;Rajashankar KR;Banerjee A
通讯作者：
Banerjee A
DOI：
10.1083/jcb.201008160
发表时间：
2010-12-27
期刊：
The Journal of cell biology
影响因子：
--
作者：
Salaun C;Greaves J;Chamberlain LH
通讯作者：
Chamberlain LH
影响因子：
4.8
作者：
Gorleku, Oforiwa A.;Barns, Anna-Marie;Chamberlain, Luke H.
通讯作者：
Chamberlain, Luke H.
影响因子：
64.8
作者：
Burr ML;Sparbier CE;Chan YC;Williamson JC;Woods K;Beavis PA;Lam EYN;Henderson MA;Bell CC;Stolzenburg S;Gilan O;Bloor S;Noori T;Morgens DW;Bassik MC;Neeson PJ;Behren A;Darcy PK;Dawson SJ;Voskoboinik I;Trapani JA;Cebon J;Lehner PJ;Dawson MA
通讯作者：
Dawson MA