PCR CLONING OF NOVEL TYROSINE PHOSPHATASES FROM RHESUS MONKEY TISSUES

酪氨酸激酶对酪氨酸磷酸化的意义 信号蛋白中的残基参与信号转导 胰岛素与胰岛素受体结合后的作用。人和 患有2型糖尿病的动物对胰岛素的作用有抵抗力。 治疗2型糖尿病的一种方法是增加胰岛素 通过抑制参与其中的酶来传递信号 去磷酸化酪氨酸(酪氨酸磷酸酶)，使其失活 信号。为了鉴定潜在的酪氨酸磷酸酶 抑制这些酶活性的药物靶点 在灵长类动物中起作用的特定磷酸酶需要被识别。 灵长类酪氨酸磷酸酶以前没有被克隆过。 目的获得胰岛素敏感组织(脂肪、肌肉和 肝脏)进行酪氨酸的聚合酶链式反应(PCR)克隆 磷酸酶。克隆是由SUGEN公司和在SUGEN，Inc.进行的。结果 组织收集在加州地区灵长类动物研究中心 中心(CRPRC)。样品取自5例正常体重和5例正常体重 肥胖的成年雄性恒河猴。脂肪(0.5g)和肌肉(0.5g)样本 在氯胺酮麻醉下通过手术活检收集。肝脏样本 (20-50毫克)通过超声引导下的穿刺针活检收集 氯胺酮麻醉。活组织检查程序由CRPRC执行 兽医。样本被放置在无菌塑料瓶中， 立即冷冻在液氮中。冰冻的组织样本是 用干冰运往加利福尼亚州红杉市的SUGEN，Inc.SUGEN HAS 从样品中克隆了几种酪氨酸磷酸酶。组织样本 也根据该项目/协议获得了用于基因阵列的 由我们实验室进行分析。未来方向没有进一步的实验 计划在这个时候，然而，有可能在未来 我们将希望从这些或其他动物身上获得更多的组织 进行类似的实验。关键词酪氨酸磷酸酶、肥胖、 糖尿病，聚合酶链式反应克隆，肌肉，脂肪，肝脏基金SUGEN，Inc. 加利福尼亚州红杉市

Significance Tyrosine kinases which phosphorylate tyrosine residues in signaling proteins are involved in transduction of insulin's actions after it binds to insulin receptors. People and animals with Type 2 diabetes are resistant to the actions of insulin. One approach to treating Type 2 diabetes is to enhance the insulin signal by inhibiting the enzymes which are involved in dephosphorylating tyrosine (Tyrosine phosphatases) which inactivates the signal. In order to identify potential tyrosine phosphatase targets for drugs that will inhibit the activity of these enzymes the specific phosphatases functioning in primates need to be identified. Primate tyrosine phosphatases have not previously been cloned. Objectives To obtain insulin sensitive tissues (fat, muscle, and liver) for polymerase chain reaction (PCR) cloning of tyrosine phosphatases. Cloning was performed by and at Sugen, Inc. Results Tissues were collected at the California Regional Primate Research Center (CRPRC). Samples were obtained from 5 normal weight and 5 obese adult male rhesus. Fat (0.5 g) and muscle (0.5 g) samples was collected by surgical biopsy under ketamine anesthesia. Liver samples (20-50 mg) were collected by ultrasound-guided needle biopsy under ketamine anesthesia. Biopsy procedures were performed by CRPRC veterinarians. Samples were placed in sterile plastic vials and immediately frozen in liquid nitrogen. The frozen tissue samples were shipped on dry ice to Sugen, Inc. in Redwood City, CA. Sugen has cloned several tyrosine phosphatases from the samples. Tissue samples have also been obtained under this project/protocol for gene array analysis by our laboratory. Future Directions No further experiments are planned at this time, however, it is possible that in the future we will want to obtain additional tissues from these or other animals for similar experiments. KEY WORDS tyrosine phosphatases, obesity, diabetes, PCR cloning, muscle, Adipose, liver FUNDING Sugen, Inc., Redwood City, CA