PHOSPHOLIPASE A1

我们之前提纯了一种可溶性磷脂酸-喜好 从牛睾丸中提取磷脂酶A1，并对其进行了克隆和序列分析。 这种酶可能在磷脂中发挥重要作用。 酸依赖的细胞信号转导，但这一作用仍有待确定。 该酶基因在人体组织中分布的研究 展示了几种反应性成分，这些成分分布在 在睾丸、脑和其他组织中的情况有所不同。研究对象： 牛酶抗体显示出交叉反应物质 在包括睾丸在内的猕猴组织中。睾丸酶可能是 从猕猴睾丸高速上清液中沉淀出来 用蛋白激酶CK2的抗体进行匀浆。此外， CK2α亚基的重组形式可以催化 天然形式和重组形式的磷酸化 磷脂酶A1和CK2α亚基的重组形式 而磷脂酶可以形成一个紧密结合的复合体x。 结果提出了CK2可能以某种方式调节的新可能性 磷脂酶的功能，并试图测试这一点 可能性目前正在进行中。此外，现场 杂交和免疫细胞化学研究计划检查 与CK2和CK2对应的mRNAs和蛋白质 磷脂酶可能是共表达的。潜力的又一发现 有趣的是，这种物质可以与人的抗体发生交叉反应 附睾尾精子中存在磷脂酶。 这种材料的相对分子质量较低(约30 Kda)，并与尾巴紧密关联。我们目前正在 试图分离出足以进行测序的这种材料的数量 用质谱仪进行分析。资助美国国立卫生研究院拨款RR00166和 霍华德·休斯医学院。霍伦巴克，D.和Glomset，J.A. 一种牛硬脂酰化转酰基酶的相互关系 睾丸膜和共同纯化的酰基转移酶。生物化学37 363-376,1998。Higgs，H.N.，han，M.H.，Johnson，G.E.和Glomset， 一种嗜磷脂酸的磷脂酶A1的克隆 牛睾丸。J.Biol.化学。273：5468-5477,1998。[此项目 在神经科学核心部分也有报道。]

We previously purified a soluble phosphatidic acid-preferring phospholipase A1 from bovine testis and cloned and sequenced its cDNA. This enzyme could potentially play an important role in phosphatidic acid-dependent cell signaling, but this role remains to be defined. Studies of the distribution of the enzyme's mRNA in human tissues demonstrated several reactive components, which were distributed differently in the testis, brain, and other tissues. Studies with antibodies to the bovine enzyme demonstrated cross-reactive material in macaque tissues including the testis. The testis enzyme could be precipitated from the high-speed supernatant of macaque testis homogenates with antibodies to the protein kinase, CK2. Furthermore, a recombinant form of the alpha subunit of CK2 could catalyze the phosphorylation of both native and recombinant forms of the phospholipase A1; and recombinant forms of the alpha subunit of CK2 and the phospholipase could form a tightly associa ted comple x. These results raise the novel possibility that CK2 may in some way regulate the function of the phospholipase, and attempts to test this possibility are currently underway. In addition, in situ hybridization and immunocytochemical studies are planned to examine the possibility that the mRNAs and proteins that correspond to CK2 and the phospholipase may be coexpressed. Another finding of potential interest is that material that cross-reacts with antibodies to the phospholipase is present in spermatozoa from the cauda epididymis. This material is of relatively low molecular mass (approximately 30 KDa) and is tightly associated with the tail. We are currently attempting to isolate amounts of this material sufficient for sequence analysis by mass spectroscopy. FUNDING NIH grant RR00166 and the Howard Hughes Medical Institute. Hollenback, D. and Glomset, J.A. On the relation between a stearoyl-specific transacylase from bovine testis membranes and a co-purifying acyltransferase. Biochemistry 37 363-376, 1998. Higgs, H.N., Han, M.H., Johnson, G.E., and Glomset, J.A. Cloning of a phosphatidic acid-preferring phospholipase A1 from bovine testis. J. Biol. Chem. 273:5468-5477, 1998. [This project was also reported in the Neuroscience Core section.]