HSP47: a tissue-specific, transformation-sensitive, collagen-binding heat shock protein of chicken embryo fibroblasts
HSP47: a tissue-specific, transformation-sensitive, collagen-binding heat shock protein of chicken embryo fibroblasts
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DOI：
10.1128/mcb.11.8.4036-4044.1991
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发表时间：
1991-08
影响因子：
5.3
通讯作者：
K. Hirayoshi;H. Kudo;H. Takechi;A. Nakai;A. Iwamatsu;K. Yamada;K. Nagata
中科院分区：
文献类型：
--
作者：
K. Hirayoshi;H. Kudo;H. Takechi;A. Nakai;A. Iwamatsu;K. Yamada;K. Nagata
我们报告的分离和表征的cDNA克隆编码热休克蛋白47，转化敏感的热休克蛋白，结合胶原蛋白。从热休克鸡胚成纤维细胞中分离的总RNA制备cDNA文库，并通过使用基于生化纯化的HSP 47的N-末端氨基酸序列制备的寡核苷酸混合物进行筛选。该cDNA插入片段全长3, 278 bp，编码一个15个氨基酸的信号肽和一个由390个氨基酸组成的成熟蛋白编码区，还包括部分5'端非编码区和一个较长的3'端非编码区。推导的氨基酸序列揭示了在C端的RDEL序列，这是一个变体的KDEL保留信号的蛋白质在内质网中的保留。北方（RNA）印迹分析和核连续试验证实，热休克对HSP 47的诱导以及劳斯肉瘤病毒转化鸡胚成纤维细胞后对其的抑制是在转录水平上调节的。同源性搜索显示，该蛋白属于丝氨酸蛋白酶抑制剂家族，即血浆丝氨酸蛋白酶抑制剂的超家族。虽然在结构上与丝氨酸蛋白酶抑制剂同源，但HSP 47缺乏被认为是抑制蛋白酶所必需的活性位点，并且似乎不与细胞内蛋白酶结合。热休克蛋白47是第一个被发现的热休克蛋白丝氨酸蛋白酶抑制剂超家族成员。相反，它是第一个不从细胞分泌的丝氨酸蛋白酶抑制剂家族成员，这可以通过在进化过程中获得RDEL保留信号来解释。
We report the isolation and characterization of a cDNA clone encoding HSP47, a transformation-sensitive heat shock protein that binds to collagen. A cDNA library was prepared from total RNA isolated from heat-shocked chicken embryo fibroblasts and screened by using oligonucleotide mixtures prepared on the basis of the N-terminal amino acid sequence of biochemically purified HSP47. The cDNA insert contained 3,278 bp, which encoded a 15-amino-acid signal peptide and a mature protein coding region consisting of 390 amino acid residues; it also included part of the 5' noncoding region and a long 3' noncoding region. The deduced amino acid sequence revealed an RDEL sequence at the C terminus, which is a variant of the KDEL retention signal for retention of proteins in the endoplasmic reticulum. Northern (RNA) blot analyses and nuclear run-on assays established that the induction of HSP47 by heat shock and its suppression after transformation of chicken embryo fibroblasts by Rous sarcoma virus are regulated at the transcriptional level. A homology search revealed that this protein belongs to the serpin family, the superfamily of plasma serine protease inhibitors. Although structurally homologous to the serpins, HSP47 lacks the active site thought to be essential for the inhibition of proteases and does not appear to bind to intracellular proteases. HSP47 is the first heat shock protein found to be a member of the serpin superfamily. Conversely, it is the first serpin family member that is not secreted from cells, which could be explained by acquisition of the RDEL retention signal during evolution.