The yeast DHHC cysteine-rich domain protein Akr1p is a palmitoyl transferase.
The yeast DHHC cysteine-rich domain protein Akr1p is a palmitoyl transferase.
复制标题
DOI：
10.1083/jcb.200206120
复制
发表时间：
2002-10-14
期刊：
影响因子：
--
通讯作者：
Davis NG
中科院分区：
文献类型：
--
作者：
Roth AF;Feng Y;Chen L;Davis NG
蛋白质棕榈酰化长期以来一直被认为是将蛋白质束缚在膜上的作用，但负责这种修饰的酶却无法识别。在此，体内和体外实验证明，Akr 1 p，一种含有DHHC富含半胱氨酸结构域（CRD）的多位膜蛋白，是棕榈酰转移酶（PTase）。在体内，我们发现酪蛋白激酶Yck 2 p是棕榈酰化的，Akr 1 p功能是这种修饰所必需的。Akr 1 p，纯化到接近同质的酵母膜，催化Yck 2 p棕榈酰化在体外，表明Akr 1 p本身是一个PTase。棕榈酰化被添加的ATP刺激。此外，在反应过程中，Akr 1 p本身是棕榈酰化的，这表明棕榈酰-Akr 1 p中间体在整个反应机制中的作用。引入Akr 1 p DHHC-CRD的突变消除了反式和自棕榈酰化活性，表明该保守序列在酶促反应中的中心参与。最后，我们的研究结果表明，棕榈酰化酵母细胞内的多种PTase特异性控制。保守的DHHC-CRD序列，我们建议，是一个进化广泛的PTase家族的签名功能。
Protein palmitoylation has been long appreciated for its role in tethering proteins to membranes, yet the enzymes responsible for this modification have eluded identification. Here, experiments in vivo and in vitro demonstrate that Akr1p, a polytopic membrane protein containing a DHHC cysteine-rich domain (CRD), is a palmitoyl transferase (PTase). In vivo, we find that the casein kinase Yck2p is palmitoylated and that Akr1p function is required for this modification. Akr1p, purified to near homogeneity from yeast membranes, catalyzes Yck2p palmitoylation in vitro, indicating that Akr1p is itself a PTase. Palmitoylation is stimulated by added ATP. Furthermore, during the reaction, Akr1p is itself palmitoylated, suggesting a role for a palmitoyl-Akr1p intermediate in the overall reaction mechanism. Mutations introduced into the Akr1p DHHC-CRD eliminate both the trans- and autopalmitoylation activities, indicating a central participation of this conserved sequence in the enzymatic reaction. Finally, our results indicate that palmitoylation within the yeast cell is controlled by multiple PTase specificities. The conserved DHHC-CRD sequence, we propose, is the signature feature of an evolutionarily widespread PTase family.
登录
查看更多内容
影响因子：
11.4
作者：
FUJIYAMA, A;MATSUMOTO, K;TAMANOI, F
通讯作者：
TAMANOI, F
影响因子：
4.5
作者：
Chen, LY;Davis, NG
通讯作者：
Davis, NG
影响因子：
5.3
作者：
DESCHENES, RJ;BROACH, JR
通讯作者：
BROACH, JR
影响因子：
4.8
作者：
BERTHIAUME, L;RESH, MD
通讯作者：
RESH, MD
影响因子：
4.3
作者：
Putilina, T;Wong, P;Gentleman, S
通讯作者：
Gentleman, S