Cloning and recombinant expression of phospholipase A2 present in rheumatoid arthritic synovial fluid.
Cloning and recombinant expression of phospholipase A2 present in rheumatoid arthritic synovial fluid.
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DOI：
10.1016/s0021-9258(18)83549-9
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发表时间：
1989-04
期刊：
影响因子：
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通讯作者：
J. Seilhamer;Waldemar Pruzanski;P. Vadas;S. Plant;J. Miller;J. Kloss;Lorin K. Johnson
中科院分区：
文献类型：
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作者：
J. Seilhamer;Waldemar Pruzanski;P. Vadas;S. Plant;J. Miller;J. Kloss;Lorin K. Johnson
来自关节炎患者的滑液含有多种形式的磷脂酶A2(PLA2)，可通过高效液相色谱法测定(Seilhamer， J.J, Plant， S., Pruzanski， W., Schilling， J., Stefanski， E., Vadas， P., and Johnson， L. K.（1989）。物化学。（东京），提交出版)。在这里，我们描述了一个人类4.5千碱基基因和800碱基对cDNA的克隆，编码了代表主要活性峰和蛋白质质量（峰a）的形式。克隆编码一个由124个氨基酸组成的成熟肽，其后是一个由20个残基组成的前肽。推导出的氨基酸序列构成了PLA2s的“II型”类酶，与其他哺乳动物来源的PLA2s相似。这是首次报道的全长哺乳动物非胰腺pla2序列。在两种不同的炎症细胞源中检测到pla2基因的活性转录。重组人峰A pla2在痘苗中作为分泌蛋白表达，在条件培养基中积累。
Synovial fluid from arthritic patients contains multiple forms of phospholipase A2(PLA2), as resolved by high performance liquid chromatography (Seilhamer, J.J., Plant, S., Pruzanski, W., Schilling, J., Stefanski, E., Vadas, P., and Johnson, L. K. (1989)J. Biochem. (Tokyo), submitted for publication). Here we describe the cloning of a human 4.5-kilobase gene and 800-base pair cDNA encoding the form representing the major peak of activity and protein mass (peak A). The clones encode a mature peptide of 124 amino acids, which follows a prepeptide of 20 residues. The deduced amino acid sequence constitutes an enzyme of the “Type II” class of PLA2s, and resembles PLA2sfrom other mammalian sources. This represents the first report of a full length mammalian non-pancreatic PLA2sequence. Active transcription of this PLA2gene was detected in two different inflammatory cell sources. Recombinant human peak A PLA2was expressed in vaccinia as a secreted protein which accumulated in conditioned medium.