SVOP is a nucleotide binding protein.
SVOP is a nucleotide binding protein.
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DOI：
10.1371/journal.pone.0005315
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Bajjalieh SM
中科院分区：
文献类型：
--
作者：
Yao J;Bajjalieh SM
突触小泡蛋白 2 (SV2) 和 SV2 相关蛋白 (SVOP) 是转运蛋白样蛋白，定位于含有神经递质的囊泡。这两种蛋白与小分子转运蛋白的主要促进子 (MF) 家族具有结构相似性。我们最近报道了 SV2 结合核苷酸，另一个 MF 家族成员人类葡萄糖转运蛋白 1 (Glut1) 也有报道这一特征。就 Glut1 而言，核苷酸结合会影响转运活性。在这项研究中，我们确定了 SVOP 是否也结合核苷酸并评估了其核苷酸结合特性。我们使用光反应 ATP 类似物、8-叠氮基-ATP[γ]生物素和纯化的重组 SVOP-FLAG 融合蛋白进行了体外光亲和标记实验。我们发现SVOP是一种核苷酸结合蛋白，尽管其底物特异性和结合位点与SV2不同。在测试的核苷酸中，ATP、GTP 和 NAD 对 SVOP-FLAG 标记显示出相同水平的抑制。剂量依赖性研究表明，SVOP 对 NAD 具有最高的亲和力，而 SV2 则以相同的亲和力结合 NAD 和 ATP。结合位点的绘图揭示了跨膜结构域 9-12 的单个区域，这与 SV2A 中大细胞质结构域中的两个结合位点形成对比。 SVOP 是被发现结合核苷酸的第三个 MF 家族成员。鉴于 SVOP、SV2 和 Glut1 中的结合位点是唯一的，因此该特征似乎是单独出现的。
Synaptic Vesicle Protein 2 (SV2) and SV2-related protein (SVOP) are transporter-like proteins that localize to neurotransmitter-containing vesicles. Both proteins share structural similarity with the major facilitator (MF) family of small molecule transporters. We recently reported that SV2 binds nucleotides, a feature that has also been reported for another MF family member, the human glucose transporter 1 (Glut1). In the case of Glut1, nucleotide binding affects transport activity. In this study, we determined if SVOP also binds nucleotides and assessed its nucleotide binding properties. We performed in vitro photoaffinity labeling experiments with the photoreactive ATP analogue, 8-azido-ATP[γ] biotin and purified recombinant SVOP-FLAG fusion protein. We found that SVOP is a nucleotide-binding protein, although both its substrate specificity and binding site differ from that of SV2. Within the nucleotides tested, ATP, GTP and NAD show same level of inhibition on SVOP-FLAG labeling. Dose dependent studies indicated that SVOP demonstrates the highest affinity for NAD, in contrast to SV2, which binds both NAD and ATP with equal affinity. Mapping of the binding site revealed a single region spanning transmembrane domains 9–12, which contrasts to the two binding sites in the large cytoplasmic domains in SV2A. SVOP is the third MF family member to be found to bind nucleotides. Given that the binding sites are unique in SVOP, SV2 and Glut1, this feature appears to have arisen separately.
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DOI：
10.1083/jcb.106.1.51
发表时间：
1988-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lowe AW;Madeddu L;Kelly RB
通讯作者：
Kelly RB
影响因子：
56.9
作者：
BAJJALIEH, SM;PETERSON, K;SCHELLER, RH
通讯作者：
SCHELLER, RH
影响因子：
5
作者：
Gillard, M;Chatelain, P;Fuks, B
通讯作者：
Fuks, B
影响因子：
4.4
作者：
Jacobsson, Josefin A.;Haitina, Tatjana;Fredriksson, Robert
通讯作者：
Fredriksson, Robert
影响因子：
3.5
作者：
Schivell, AE;Mochida, S;Bajjalieh, SM
通讯作者：
Bajjalieh, SM