Near-UV circular dichroism of band 3. Evidence for intradomain conformational changes and interdomain interactions.
Near-UV circular dichroism of band 3. Evidence for intradomain conformational changes and interdomain interactions.
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谱带 3 的近紫外圆二色性。域内构象变化和域间相互作用的证据。
DOI：
10.1021/bi00053a035
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Salhany,JM
中科院分区：
文献类型：
--
作者：
Batenjany,MM;Mizukami,H;Salhany,JM
1992年10月20日收到的修订版Mandarin pt摘要：近紫外圆二色性（CD）用于鉴定人红细胞带3（氯化物/碳酸氢盐交换蛋白）三级结构的差异，这是阴离子转运抑制剂与单体内二苯乙烯二磺酸盐（ISD）位点共价结合的结果。将分离的完整带3与其膜结构域（B3 MD）进行比较。观察到光谱差异，其中涉及域内效应，因为它们被视为与完整的带3和B3 MD，或域间效应，因为它们被观察到只有B3 MD，但被抑制时，细胞质域连接。域内效应涉及在低于280 nm的光谱的Phe/Tyr区域中的光学活性的显著损失。只有当ISD位点在pH 7.4时具有共价结合的芪二磺酸盐时才能看到。在加合物形成后将pH升高至9.6，不可逆地“正常化”了这种光谱变化。在292 nm处的Trp光谱区域中鉴定了畴间效应。当大的共价配体如DIDS（4，4 '-diisothiocyanatostilbene-2，2 H-disulfonate）与B3 MD结合时，在292 nm处的光学活性显著增加，但当相同的配体与完整的条带3结合时，光学活性没有显著增加。这些后者的结果提供的证据表明，某些方面的整体结构域的构象反应被抑制的存在下，一个附加的胞质结构域。带3是一种完整的膜蛋白，它促进氯离子交换为碳酸氢盐穿过红细胞膜，也为某些细胞骨架和胞质蛋白提供结合位点（Passow，1986; Salhany，1990）。这种糖蛋白（Mr 95 000）以二聚体和四聚体的混合物原位存在（Salhany等人，1990;凯西和Reithmeier，1991），并且由两个域组成。N-末端胞质结构域（Mi 40 000）延伸到细胞中并提供蛋白质结合位点（Low，1986）。C-末端整合结构域（Mr 55 000）跨越双层，并且可以在没有连接的细胞质结构域的帮助下转运阴离子（Lepke & Passow，1976; Grinstein等人，1978; Lepke等人，1992年）。它是蛋白质的组成结构域，含有一大类阴离子交换抑制剂的结合位点（Passow，1986; Salhany，1990; Cabantchik & Greger，1992）。关于带3结构/功能的主要观点是，在整合域中存在一个转运位点，该转运位点可以一次结合一个阴离子，并以一种独立于原位存在的各种寡聚体形式中相邻亚基的功能的方式在面向内和面向外的状态之间交替构象
Revised Manuscript Received October 20, 1992 abstract: Near-UV circular dichroism (CD) was used to identify differences in the tertiary structure of human erythrocyte band 3, the chloride/bicarbonate exchange protein, consequent to covalent binding of anion transportinhibitors to the intramonomeric stilbenedisulfonate (ISD) site. Isolatedintact band 3 and its membrane domain (B3MD) were compared. Spectral differences were observed which involved intradomain effects, in that they were seen both with intact band 3 and with B3MD, or interdomain effects, in that they were observed only for B3MD, but were inhibited when the cytoplasmic domain was attached. The intradomain effect involved a significant loss in optical activity in the Phe/Tyr region of the spectrum below 280 nm. It was seen only when the ISD site had stilbenedisulfonates bound covalently at pH 7.4. Raising the pH to 9.6 after adduct formation “normalized” this spectral change irreversibly. The interdomain effect was identified in the Trp spectral region at 292 nm. There was a significant increase inoptical activity at 292 nm when bulky covalent ligandssuch as DIDS (4, 4'-diisothiocyanatostilbene-2, 2/-disulfonate) were bound to B3MD, but not when the same ligands were bound to intact band 3. These latter results offer evidence that certain aspects of the conformational response of the integral domain are inhibited bythe presence of an attached cytoplasmic domain. The potential significance of interdomain interactions to band 3 function is discussed briefly.Band 3 is an integral membrane protein which facilitates the exchange of chloride for bicarbonate across the erythrocyte membrane and also provides binding sites for certaincytoskeletal and cytosolic proteins (Passow, 1986; Salhany, 1990). This glycoprotein (Mr 95 000) exists as a mixture of dimers and tetramers in situ (Salhany et al., 1990; Casey & Reithmeier, 1991) and is composed of two domains. The N-terminal cytoplasmic domain {Mi 40 000) extends into the cell and provides the protein binding sites (Low, 1986). The C-terminal integral domain {Mr 55 000) spans the bilayer and can transport anions without the aid of an attached cytoplasmic domain (Lepke & Passow, 1976; Grinstein et al., 1978; Lepke et al., 1992). It is the integral domain of the protein which contains binding sites for a large class of inhibitors of anion exchange (Passow, 1986; Salhany, 1990; Cabantchik & Greger, 1992). The predominant view of band 3 structure/function has been that a transport siteexists within the integral domain which can bind one anion at a time and alternate conformation between inside-facing and outside-facing states in a manner which is independent of the functioning of neighboring subunits within the various oligomeric forms which exist in situ
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红细胞带 3 蛋白的三个区域在酪氨酸上被磷酸化：通过固相测序结合毛细管电泳来表征磷酸化位点。
DOI：
--
发表时间：
1991
期刊：
Biochimica et Biophysica Acta
影响因子：
--
作者：
D. Yannoukakos;Helmut E. Meyer;Corinne Vasseur;Catherine Driancourt;H. Wajcman;Elizabeth Bursaux
通讯作者：
Elizabeth Bursaux
人带 3 的赖氨酸 539 对于离子传输或二苯乙烯二磺酸盐的抑制不是必需的。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Garcia,AM;Lodish,HF
通讯作者：
Lodish,HF
与红细胞膜结合的血红蛋白的光散射测量。
影响因子：
2.9
作者：
J. M. Salhany;K. A. Cordes;E. Gaines
通讯作者：
E. Gaines
DOI：
10.1073/pnas.86.23.9089
发表时间：
1989-12-01
影响因子：
11.1
作者：
LUX, SE;JOHN, KM;LODISH, HF
通讯作者：
LODISH, HF
通过分析超速离心分析膜蛋白之间的复杂关联：红细胞膜蛋白带 3 和锚蛋白的研究
DOI：
--
发表时间：
1990
期刊：
影响因子：
--
作者：
K. Mulzer;L. Kampmann;P. Petrasch;D. Schubert
通讯作者：
D. Schubert