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来自巴西橡胶树的一个编码小橡胶粒子蛋白的新型cDNA克隆的分离、鉴定及功能分析。

Isolation, characterization, and functional analysis of a novel cDNA clone encoding a small rubber particle protein from Hevea brasiliensis.

作者信息

Oh S K, Kang H, Shin D H, Yang J, Chow K S, Yeang H Y, Wagner B, Breiteneder H, Han K H

机构信息

Kumho Life and Environmental Science Laboratory, 1 Oryong-Dong, Puk-Gu, Kwangju 500-480, Korea.

出版信息

J Biol Chem. 1999 Jun 11;274(24):17132-8. doi: 10.1074/jbc.274.24.17132.

Abstract

Biochemical evidence reported so far suggests that rubber synthesis takes place on the surface of rubber particles suspended in the latex of Hevea brasiliensis. We have isolated and characterized a cDNA clone that encodes a protein tightly bound on a small rubber particle. We named this protein small rubber particle protein (SRPP). Prior to this study, this protein was known as a latex allergen, and only its partial amino acid sequence was reported. Sequence analysis revealed that this protein is highly homologous to the rubber elongation factor and the Phaseolus vulgaris stress-related protein. Southern and Northern analyses indicate that the protein is encoded by a single gene and highly expressed in latex. An allergenicity test using the recombinant protein confirmed that the cloned cDNA encodes the known 24-kDa latex allergen. Neither ethylene stimulation nor wounding changed the transcript level of the SRPP gene in H. brasiliensis. An in vitro rubber assay showed that the protein plays a positive role in rubber biosynthesis. Therefore, it is likely that SRPP is a part of the rubber biosynthesis machinery, if not the rubber polymerase, along with the rubber elongation factor.

摘要

目前报道的生化证据表明,橡胶合成发生在巴西橡胶树胶乳中悬浮的橡胶粒子表面。我们分离并鉴定了一个cDNA克隆,它编码一种紧密结合在小橡胶粒子上的蛋白质。我们将这种蛋白质命名为小橡胶粒子蛋白(SRPP)。在本研究之前,这种蛋白质被认为是一种胶乳过敏原,并且仅报道了其部分氨基酸序列。序列分析表明,该蛋白质与橡胶延长因子和菜豆应激相关蛋白高度同源。Southern和Northern分析表明,该蛋白质由单一基因编码,并在胶乳中高表达。使用重组蛋白进行的过敏原性测试证实,克隆的cDNA编码已知的24 kDa胶乳过敏原。乙烯刺激和创伤均未改变巴西橡胶树中SRPP基因的转录水平。体外橡胶测定表明,该蛋白质在橡胶生物合成中起积极作用。因此,SRPP很可能是橡胶生物合成机制的一部分,即便不是橡胶聚合酶,也是与橡胶延长因子一起的一部分。

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