1H, 13C and 15N resonance assignments of the VAP-A: OSBP complex
1H, 13C and 15N resonance assignments of the VAP-A: OSBP complex
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VAP-A 的 1H、13C 和 15N 共振分配：OSBP 复合物
DOI：
10.1007/s10858-006-9057-2
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发表时间：
2006
影响因子：
2.7
通讯作者：
C. Kojima
中科院分区：
文献类型：
--
作者：
Kyoko Furuita;M. Mishima;C. Kojima
氧固醇结合蛋白（OSBP）通过与VAMP相关蛋白A（VAP-A）形成复合物来介导25-羟基胆固醇（25-OHC）的运输。VAP-A是一种内质网（ER）整合膜蛋白，含有一个细胞质主要精子蛋白（MSP）结构域。OSBP中存在一个FFAT（指酸性区中的两个苯丙氨酸[FF]）基序，该基序被VAP-A的MSP结构域识别。因此，相互作用促进OSBP定位于ER，其中新合成的25-羟基胆固醇（25-OHC）有效地结合OSBP（Wyles等人，2002年）。作为理解VAP-A和OSBP介导的25-OHC运输机制的第一步，我们对由VAP-A MSP结构域（5-128）和含有FFAT基序的OSBP肽（345-379）组成的复合物进行了NMR研究。对13 C和15 N标记的VAP-A与未标记的OSBP、13 C和15 N标记的OSBP与未标记的VAP-A以及15 N标记的VAP-A与15 N标记的OSBP组成的复合物进行了2D、3D和4D NMR实验。除VAP-A的N34 C′外，其余骨架核（1HN，15 N，13 Co和13 C ′）的共振峰均已归属。此外，超过90%的侧链15 H和13 C共振的配合物也被指定。OSBP的T367-G373显示可能源自次要构象的次要峰。BMRB存款，保藏号7025。参考文献：Wyles et al.（2002）J. Biol. Chem.，277，29908-29918。
The 25-hydroxycholesterol (25-OHC) traffic by oxysterol binding protein (OSBP) is mediated by complex formation of OSBP with VAMP-associated protein-A (VAP-A). VAP-A is an endoplasmic reticulum (ER) integral membrane protein that contains a cytoplasmic major sperm protein (MSP) domain. There is a FFAT (referring to two phenylalanines [FF] in an acidic tract) motif in OSBP that is recognized by the MSP domain of VAP-A. Thus, the interaction facilitates localization of OSBP to the ER, where newly synthesized 25-hydroxycholesterol (25-OHC) efficiently binds to OSBP (Wyles et al., 2002). As a first step toward understanding the mechanism of 25-OHC traffic mediated by VAP-A and OSBP, we performed NMR studies of the complex composed of a VAP-A MSP domain (5–128) and an OSBP peptide (345–379) containing the FFAT motif. 2D, 3D and 4D NMR experiments were performed with the complex composed of 13C-and 15N-labeled VAP-A and non-labeled OSBP, 13C-and 15N-labeled OSBP and non-labeled VAP-A, and 15N-labeled VAP-A and 15N-labeled OSBP. All resonances of the backbone nuclei (1HN, 15N, 13Co and 13C′) of the complex were assigned with the exception of N34 C′ of VAP-A. Furthermore, more than 90% of the side chain 15H and 13C resonances of the complex were also assigned. T367-G373 of OSBP showed minor peaks possibly derived from a minor conformation. BMRB deposit with accession No. 7025. Reference: Wyles et al.(2002) J. Biol. Chem., 277, 29908–29918.