Harvey T J, Hooper J D, Myers S A, Stephenson S A, Ashworth L K, Clements J A
Centre for Molecular Biotechnology, School of Life Sciences, Queensland University of Technology, Brisbane, Queensland 4001, Australia.
J Biol Chem. 2000 Dec 1;275(48):37397-406. doi: 10.1074/jbc.M004525200.
The tissue or glandular kallikreins (KLK) are members of a highly conserved multigene family encoding serine proteases that are central to many biological processes. The rodent KLK families are large, highly conserved and clustered at one locus. The human KLK gene family is clustered on chromosome 19q13.3-13.4, and until recently consisted of just three members. However, recent studies have identified up to 11 new members of the KLK family that are less conserved than their rodent counterparts. Using a Southern blot and sequence analysis of 10 BACs and cosmids spanning approximately 400 kilobases (kb) either side of the original KLK 60-kb locus, we demonstrated that these genes also lie adjacent to this. We have also clarified the position of several microsatellite markers in relation to the extended KLK locus. Moreover, from Southern blot analysis of the cosmids and BACs with a degenerate oligonucleotide probe to the histidine-encoding region of serine proteases, we have shown that there are no other serine protease genes approximately 400 kb centromeric and 220 kb telomeric of the extended locus. We performed an extensive analysis of the expression patterns of these genes by poly(A)(+) RNA dot blot and reverse transcriptase-polymerase chain reaction analysis, and demonstrated a diverse pattern of expression. Of interest are clusters of genes with high prostate (KLK2-4) and pancreatic (KLK6-13) expression suggesting evolutionary conservation of elements conferring tissue specificity. From these findings, it is likely that the human KLK gene family consists of just 14 clustered genes within 300 kb and thus is of a comparable size to the rodent families (13-24 genes within 310 and 480 kb, respectively). In contrast to the rodent families, the newest members of the human KLK family are much less conserved in sequence (23-44% at the protein level) and appear to consist of at least four subfamilies. In addition, like the rat, these genes are expressed at varying levels in a diverse range of tissues although they exhibit quite distinct patterns of expression.
组织或腺体激肽释放酶（KLK）是一个高度保守的多基因家族的成员，该家族编码丝氨酸蛋白酶，这些酶在许多生物学过程中起核心作用。啮齿动物的KLK家族庞大、高度保守且聚集在一个基因座上。人类KLK基因家族聚集在19号染色体的19q13.3 - 13.4区域，直到最近还仅由三个成员组成。然而，最近的研究已鉴定出多达11个KLK家族的新成员，它们的保守程度低于其啮齿动物对应物。通过对跨越原始KLK 60 kb基因座两侧约400千碱基（kb）的10个细菌人工染色体（BAC）和黏粒进行Southern印迹和序列分析，我们证明这些基因也位于其附近。我们还阐明了几个微卫星标记相对于扩展的KLK基因座的位置。此外，用针对丝氨酸蛋白酶组氨酸编码区的简并寡核苷酸探针，对黏粒和BAC进行Southern印迹分析，结果表明在扩展基因座着丝粒方向约400 kb和端粒方向约220 kb范围内没有其他丝氨酸蛋白酶基因。我们通过聚腺苷酸（poly(A)）(+) RNA斑点印迹和逆转录 - 聚合酶链反应分析对这些基因的表达模式进行了广泛分析，并证明了其多样的表达模式。有趣的是，具有高前列腺（KLK2 - 4）和胰腺（KLK6 - 13）表达的基因簇表明赋予组织特异性的元件具有进化保守性。基于这些发现，人类KLK基因家族可能仅由300 kb范围内的14个聚集基因组成，因此其大小与啮齿动物家族相当（分别在310 kb和480 kb范围内有13 - 24个基因）。与啮齿动物家族不同，人类KLK家族的最新成员在序列上保守性要低得多（蛋白质水平为23 - 44%），并且似乎至少由四个亚家族组成。此外，与大鼠一样，这些基因在多种组织中以不同水平表达，尽管它们表现出相当独特的表达模式。
