Cloning and expression of rat lung acidic Ca2+-independent PLA2 and its organ distribution
Cloning and expression of rat lung acidic Ca2+-independent PLA2 and its organ distribution
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DOI：
10.1152/ajplung.1998.274.5.l750
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发表时间：
1998-05-01
影响因子：
4.9
通讯作者：
Fisher, AB
中科院分区：
文献类型：
--
作者：
Kim, TS;Dodia, C;Fisher, AB
以人酸性磷脂酶A(2)为探针，从大鼠肺颗粒细胞的c DNA文库中克隆了一个大鼠酸性非钙非依赖性磷脂酶A(2)(AIPLA(2))(T.S.Kim，C.S.Sundaresh，S.I.Feinstein，C.Dodia，W.R.Skach，M.K.Jain，T.Nagase，N.Seki，K.Ishikawa，N.Nomura，A.B.Fisher)。J.Biol.化学。272：2542-2550,1997)。此外，从GenBank和DBEST数据库中的几个小鼠克隆中构建了小鼠AIPLA(2)的共同序列。每个序列编码一个224个氨基酸的蛋白质，三个物种之间的氨基酸同源性为88%，并保守一个假定的脂肪酶基序(GDSWG)。将大鼠克隆产生的mRNA在麦胚系统中翻译后，可表达出与人类酶相似的PLA2活性，即酸性最适pH和不依赖钙离子。用人cDNA探针、多克隆抗体和单抗研究了AIPLA2在大鼠组织中的定位和活性。AIPLA(2)存在于肺中，高水平的基因和蛋白表达以及酶活性被抗PLA2抗体和过渡态类似物1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol(MJ33MJ33)抑制。免疫细胞化学显示肺泡II型细胞、肺泡巨噬细胞和细支气管壁上皮细胞表达AIPLA2。在脑、心、肝、肾、脾和肠中，AIPLA2(2)的mRNA含量为
A clone for a rat acidic Ca2+-independent phospholipase A(2) (aiPLA(2)) was isolated from a cDNA library prepared from rat granular pneumocytes with a probe based on the human aiPLA(2) sequence (T. S. Kim, C. S. Sundaresh, S. I. Feinstein, C. Dodia, W. R. Skach, M. K. Jain, T. Nagase, N. Seki, K. Ishikawa, N. Nomura, and A. B. Fisher. J. Biol. Chem. 272: 2542-2550, 1997). In addition, a consensus sequence for mouse aiPLA(2) was constructed from several mouse cDNA clones in the GenBank and dbEST databases. Each sequence codes for a 224-amino acid protein with 88% identity of the amino acids among the three species and conservation of a putative lipase motif (GDSWG). Translation of mRNA produced from the rat clone in a wheat germ system resulted in expression of PLA(2) activity with properties similar to those of the human enzyme, i.e., acidic pH optimum and Ca2+ independence. The localization of aiPLA(2) in rat tissues was studied with the human cDNA probe, polyclonal and monoclonal antibodies, and aiPLA(2) activity. aiPLA(2) is present in the lung as evidenced by high levels of mRNA and protein expression and by enzymatic activity that is inhibited by anti-PLA(2) antibody and by the transition state analog 1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol (MJ33). Immunocytochemistry showed the presence of aiPLA(2) in alveolar type II cells, alveolar macrophages, and bronchiolar epithelium. In the brain, heart, liver, kidney, spleen, and intestine, aiPLA(2) mRNA content was