Phospho-Phospholamban (Ser16/Thr17) Antibody

• 产品编号：CST-8496S      品牌：CST       原厂货号：8496S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

只有当丝氨酸16位和苏氨酸17位磷酸化时，磷酸-受磷蛋白(Ser16/Thr17)抗体识别内源性的受磷蛋白（Ser16/Thr17）。该抗体不能检测到单-或非磷酸化磷蛋白。该多克隆抗体通过用合成磷酸肽免疫动物制备，该合成肽与人受磷蛋白Ser16/Thr17附近的残基一样。抗体是由蛋白A和肽亲和层析纯化。受磷蛋白(PLN)是作为内质网的一个主要磷酸化蛋白质成分鉴定出来的（1）。它的名字“lamban”，来自希腊字的“lambano”，意思是“接受”，如此命名是基于这样的事实：在响应心脏刺激时，受磷蛋白在丝氨酸和苏氨酸残基大量磷酸化（1）。尽管由于其在SDS-PAGE上的电泳迁移率，最初被认为是一个单一的20-25 kDa蛋白质，但PLN实际上是一个有52个氨基酸、6 kDa的跨膜蛋白，即使在SDS存在下，它也能形成稳定的同源寡聚体（2）。尽管受磷蛋白在心脏组织中高表达，但其在骨骼肌和平滑肌中也有所表达（3）。PLN的定位被限制在内质网，在SR上PLN可作为一个肉瘤内质网钙ATP酶，心肌肌浆网钙泵的调节器（4）。PLN直接与SERCA结合，有效降低其对钙的亲和力，从而减少钙转运到内质网。通过蛋白激酶A或强直性肌营养不良蛋白激酶在PLN丝氨酸16位的磷酸化和/或通过Ca2+/钙调蛋白依赖蛋白激酶在苏氨酸17位的磷酸化，都将导致来自肌浆网钙泵的PLN释放，减轻抑制作用，并通过内质网增强钙吸收(综述5,6)。长期以来，人们一直认为，丝氨酸16位和苏氨酸17位的磷酸化是顺序发生的，但是越来越多的证据表明，磷酸化，特别是在苏氨酸17位，可能是受到不同的调节(综述 7,8)。心脏衰竭的啮齿动物模型表明，在调节钙流和收缩性中，PLN的表达水平和磷酸化程度都是至关重要的（综述 9-11）。PLN表达的缺失或下降，促进钙流增加，增强心肌收缩，而PLN过度表达将导致肌浆网钙泵封存，减少钙流，降低收缩性，在高血压和心肌病的小鼠模型中，降低心脏功能障碍和衰竭的救援（综述10）。在人类中，已发现PLN不同的基因突变会导致PLN蛋白下降或不表达，或PLN、SERCA和/或调节蛋白之间的结合缺陷（14,15），这两个结果都会导致高血压性心肌病和心脏衰竭。有趣的是，在啮齿类中，也可以发现人类大多数PLN缺陷表型，反之亦然，也有一些在啮齿动物和人中，PLN缺陷导致不同的心脏疾病类型和严重程度，二者共同的作用机制还不清楚。

1. Kirchberber, M.A. et al. (1975) Recent Adv Stud Cardiac Struct Metab 5, 103-15.
2. Zhan, Q.Q. et al. (1991) J Biol Chem 266, 21810-4.
3. Fujii, J. et al. (1991) J Biol Chem 266, 11669-75.
4. Tada, M. and Kirchberger, M.A. Recent Adv Stud Cardiac Struct Metab 11, 265-72.
5. Traaseth, N.J. et al. (2008) Biochemistry 47, 3-13.
6. Bhupathy, P. et al. (2007) J Mol Cell Cardiol 42, 903-11.
7. Hagemann, D. and Xiao, R.P. (2002) Trends Cardiovasc Med 12, 51-6.
8. Mattiazzi, A. et al. (2005) Cardiovasc Res 68, 366-75.
9. Chu, G. and Kranias, E.G. (2006) Novartis Found Symp 274, 156-71; discussion 172-5, 272-6.
10. Schwinger, R.H. and Frank, K.F. (2003) Sci STKE 2003, pe15.
11. MacLennan, D.H. and Kranias, E.G. (2003) Nat Rev Mol Cell Biol 4, 566-77.
12. Haghighi, K. et al. (2008) Hum Mutat 29, 640-7.
13. Haghighi, K. et al. (2003) J Clin Invest 111, 869-76.
14. Schmitt, J.P. et al. (2003) Science 299, 1410-3.
15. Haghighi, K. et al. (2006) Proc Natl Acad Sci USA 103, 1388-93.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Human, Mouse, Bovine, Dog, Pig

Specificity / Sensitivity

Phospho-Phospholamban (Ser16/Thr17) Antibody recognizes endogenous levels of phospholamban protein only when phosphorylated at Ser16 and Thr17. This antibody does not detect mono- or non-phosphorylated phospholamban.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser16/Thr17 of human phospholamban protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Phospholamban (PLN) was identified as a major phosphoprotein component of the sarcoplasmic reticulum (SR) (1). Its name, "lamban", is derived from the greek word "lambano" meaning "to receive", so named due to the fact that phospholamban is heavily phosphorylated on serine and threonine residues in response to cardiac stimulation (1). Although originally thought to be a single 20-25 kDa protein due to its electrophoretic mobility on SDS-PAGE, PLN is actually a 52 amino acid, 6 kDa, membrane-spanning protein capable of forming stable homooligomers, even in the presence of SDS (2). Despite very high expression in cardiac tissue, phospholamban is also expressed in skeletal and smooth muscle (3). Localization of PLN is limited to the SR, where it serves as a regulator of the sarco-endoplasmic reticulum calcium ATPase, SERCA (4). PLN binds directly to SERCA and effectively lowers its affinity for calcium, thus reducing calcium transport into the SR. Phosphorylation of PLN at Ser16 by Protein Kinase A or myotonic dystrophy protein kinase and/or phosphorylation at Thr17 by Ca2+/calmodulin-dependent protein kinase results in release of PLN from SERCA, relief of this inhibition, and increased calcium uptake by the SR (reviewed in 5,6). It has long been held that phosphorylation at Ser16 and Thr17 occurs sequentially, but increasing evidence suggests that phosphorylation, especially at Thr17, may be differentially regulated (reviewed in 7,8). 
 
 Rodent models of heart failure have shown that the expression level and degree of phosphorylation of PLN are critical in modulating calcium flux and contractility (reviewed in 9-11). Deletion or decreased expression of PLN promotes increased calcium flux and increased cardiac contractility, whereas overexpression of PLN results in sequestration of SERCA, decreased calcium flux, reduced contractility, and rescue of cardiac dysfunction and failure in mouse models of hypertension and cardiomyopathy (reviewed in 10). Distinct mutations in PLN have been detected in humans, resulting either in decreased or no expression of PLN protein (12,13) or binding defects between PLN, SERCA and/or regulatory proteins (14,15), both of which result in cardiac myopathy and heart failure. Interestingly, while the human phenotype of most PLN defects mimic those seen in rodent and vice versa, there are some instances where the type and severity of cardiac disease resulting from PLN mutations in rodent and human differ, making a consensus mechanism elusive.

1. Kirchberber, M.A. et al. (1975) Recent Adv Stud Cardiac Struct Metab 5, 103-15.
2. Zhan, Q.Q. et al. (1991) J Biol Chem 266, 21810-4.
3. Fujii, J. et al. (1991) J Biol Chem 266, 11669-75.
4. Tada, M. and Kirchberger, M.A. Recent Adv Stud Cardiac Struct Metab 11, 265-72.
5. Traaseth, N.J. et al. (2008) Biochemistry 47, 3-13.
6. Bhupathy, P. et al. (2007) J Mol Cell Cardiol 42, 903-11.
7. Hagemann, D. and Xiao, R.P. (2002) Trends Cardiovasc Med 12, 51-6.
8. Mattiazzi, A. et al. (2005) Cardiovasc Res 68, 366-75.
9. Chu, G. and Kranias, E.G. (2006) Novartis Found Symp 274, 156-71; discussion 172-5, 272-6.
10. Schwinger, R.H. and Frank, K.F. (2003) Sci STKE 2003, pe15.
11. MacLennan, D.H. and Kranias, E.G. (2003) Nat Rev Mol Cell Biol 4, 566-77.
12. Haghighi, K. et al. (2008) Hum Mutat 29, 640-7.
13. Haghighi, K. et al. (2003) J Clin Invest 111, 869-76.
14. Schmitt, J.P. et al. (2003) Science 299, 1410-3.
15. Haghighi, K. et al. (2006) Proc Natl Acad Sci USA 103, 1388-93.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C.
Do not aliquot the antibody