Identification of a new protein that binds Src and PKC

结合 Src 和 PKC 的新蛋白的鉴定

• 批准号: 6508827
• 负责人: Stephen Paul Soltoff
• 金额: $ 16.62万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6508827
• 关键词: binding proteins; immunofluorescence technique; laboratory rabbit; laboratory rat; mass spectrometry; membrane proteins; molecular biology information system; protein binding; protein kinase C; protein localization; protein sequence; protein structure function; protooncogene; salivary glands

DESCRIPTION (provided by applicant): The broad long-term objective of this application is to determine the function of a newly identified protein that may serve as a scaffolding protein or adapter molecule for Protein Kinase C-delta (PKCdelta) and Src, two important signaling molecules that may contribute to salivary and epithelial functions. Our hypothesis is that we have identified a protein that is the product of a newly identified novel gene, and we propose a model to explain how PKCdelta and Src interact with this protein. If verified, this will serve as a basis upon which future research can be conducted. Salivary gland cells function to secrete fluids and proteins that are important to the health of the oral cavity. The proper biological activity of these cells depends on multiple proteins, including kinases that phosphorylate their appropriate substrates. One way that kinases associate with their substrates is through scaffolding proteins. Protein kinases C plays important roles in biological events in salivary glands, and understanding how PKC activities are controlled and directed will aid our understanding of salivary function, which is compromised in Sjgren's Syndrome and other situations. In salivary cells, PKCdelta is phosphorylated on tyrosine residues in response to neurotransmitters and conditions that alter fluid secretion and the permeability of tight junctions. The tyrosine phosphorylation of PKCdelta is mediated by Src or Src-related tyrosine kinases. We posit that a protein that we call "p130" is a transmembrane protein that represents a new type of anchoring protein for PKCdelta and Src. It is likely to play an important role in multiple types of epithelia. Using a Mass Spectrometry approach, we identified p130 by searching sequences in public databases. A recent report suggests that a newly identified gene encodes p130. The Specific Aims include: (1) To characterize the functional interrelationships among PKCdelta, Src, and p130; (2) To expand our studies of p130 in multiple types of salivary cells, including parotid and submandibular cells; and (3) To generate antibodies to p130 and identify and localize endogenous p130 within cells. These Aims should be achievable within the 2-year timetable of this project and allow us to determine the veracity of our hypothesis. Future efforts will determine if this new protein alters cell morphology, tight junction formation, or other functions in salivary cells.

描述（由申请人提供）：本应用的广泛长期目标是确定新近鉴定的蛋白质的功能，该蛋白可以用作蛋白激酶C-DELTA（PKCDELTA）和SRC的脚手架蛋白质或衔接子分子，这是两个重要的信号分子，可能有助于溶性和上皮功能。我们的假设是，我们已经确定了一种蛋白质，该蛋白质是新鉴定的新基因的产物，我们提出了一个模型来解释PKCDELTA和SRC如何与该蛋白质相互作用。如果经过验证，这将作为可以进行未来研究的基础。唾液腺细胞发挥作用，分泌对口腔健康重要的液体和蛋白质。这些细胞的适当生物学活性取决于多种蛋白质，包括磷酸化其适当底物的激酶。激酶与底物相关的一种方法是通过脚手架蛋白。蛋白激酶C在唾液腺中的生物事件中起重要作用，并且了解如何控制PKC活动将有助于我们对唾液功能的理解，唾液功能在SJ Gren综合征和其他情况下受到损害。在唾液细胞中，PKCDELTA响应于神经递质和疾病的酪氨酸残基上磷酸化，并改变了液体分泌和紧密连接的渗透性。 PKCDELTA的酪氨酸磷酸化是由SRC或与SRC相关的酪氨酸激酶介导的。我们认为我们称为“ P130”的蛋白质是一种跨膜蛋白，代表了PKCDELTA和SRC的一种新型锚定蛋白。它可能在多种类型的上皮中发挥重要作用。使用质谱方法，我们通过在公共数据库中搜索序列来确定P130。最近的一份报告表明，新鉴定的基因编码P130。具体目的包括：（1）表征PKCDELTA，SRC和P130之间的功能相互关系； （2）扩大我们对多种类型的唾液细胞的P130研究，包括腮腺和下颌细胞； （3）生成对P130的抗体，并在细胞内识别和定位内源性P130。这些目标应在该项目的两年时间表中实现，并允许我们确定假设的真实性。未来的努力将确定这种新蛋白质是否改变了细胞形态，紧密结的形成或唾液细胞中的其他功能。