Seidah N G, Gaspar L, Mion P, Marcinkiewicz M, Mbikay M, Chrétien M
J.A. DeSève Laboratory of Biochemical Neuroendocrinology, Clinical Research Institute of Montreal, Quebec, Canada.
DNA Cell Biol. 1990 Jul-Aug;9(6):415-24. doi: 10.1089/dna.1990.9.415.
Based on the concept of sequence conservation around the active sites of serine proteinases, polymerase chain reaction applied to mRNA amplification allowed us to obtain a 260-bp probe which was used to screen a mouse pituitary cDNA library. The primers used derived from the cDNA sequence of active sites Ser* and Asn* of human furin. Two cDNA sequences were obtained from a number of positive clones. These code for two similar but distinct structures (mPC1 and mPC2), each being homologous to yeast Kex2 and human furin. In situ hybridization (mPC1) and Northern blots (mPC1 = 3.0 kb and mPC2 = 2.8 and 4.8 kb) demonstrated tissue and cellular specificity of expression, only within endocrine and neuroendocrine cells. These data suggest that mPC1 and mPC2 represent prime candidates for tissue-specific pro-hormone converting proteinases.
基于丝氨酸蛋白酶活性位点周围序列保守性的概念，应用于mRNA扩增的聚合酶链反应使我们获得了一个260 bp的探针，该探针用于筛选小鼠垂体cDNA文库。所用引物源自人弗林蛋白酶活性位点Ser和Asn的cDNA序列。从多个阳性克隆中获得了两个cDNA序列。它们编码两种相似但不同的结构（mPC1和mPC2），每种结构都与酵母Kex2和人弗林蛋白酶同源。原位杂交（mPC1）和Northern印迹分析（mPC1 = 3.0 kb，mPC2 = 2.8和4.8 kb）证明了表达的组织和细胞特异性，仅在内分泌和神经内分泌细胞中表达。这些数据表明，mPC1和mPC2是组织特异性激素原转化蛋白酶的主要候选者。
