Characterization of physiological function of the plasma proteins, PFBP and IHRP with knockout mice.

1)PHBP-PHBP（Plasma Hyronan-Binding Protein）是本课题组于1996年从人血浆中发现的一种蛋白质，含有三个EGF、一个Kringle和一个丝氨酸蛋白酶结构域，当一个特定的肽键被切割时，PHBP被激活并显示出蛋白水解活性。为了阐明其功能，我们尝试制造PHBP基因敲除小鼠。在确定小鼠PHBP基因的基因组结构后，构建了靶向载体，以新霉素耐受基因取代第一外显子的1 kb区域。2）IHRPIHRP（Inter a trypsin inhibitor Heavy chain Related Protein）是本课题组于1996年从人血浆中发现的一种胰蛋白酶抑制剂重链相关蛋白，其氨基酸序列与ITT重链有70%的同源性，但IHRP不具有ITT所具有的胰蛋白酶抑制活性。由于THRP的功能尚未阐明，我们还尝试了IHRP基因敲除小鼠的制备。目前我们已经确定了小鼠IHRP基因的结构以及小鼠IHRP mRNA的核苷酸序列。3）GBP 28的研究在我们研究PHBP和IHRP的同时，我们还从人血浆中发现了GBP 28。GBP 28（28 kd的明胶结合蛋白）是Matsuzawa等（Osaka univ.）发现的脂肪细胞特异性mRNA的apM 1 mRNA的翻译产物。Matsuzawa后来将这种蛋白质命名为脂联素。2001年发现GBP 28/adiponectin有可能作为抗糖尿病药物。目前，我们也在尝试生产IHRP基因敲除小鼠。

1) Studies on PHBPPHBP (Plasma Hyaluronan-Binding Protein)was found from human blood plasma by our group in 1996.PHBP contains three EGFs, a Kringle and a serine protease domains, and is activated to reveal proteolytic activity when a specific peptide bond is cleaved. To elucidate its functions, we tried to produce PHBP gene knockout mice. After the genomic structure of mouse PHBP gene was determined, a targeting vector was constructed to substitute a neomycin tolerant gene for a 1 kb region of the first exon. Since an ES cell clone which was knockout with PHBP gene was produced, we are trying to produce the knockout mice.2) Studies on IHRPIHRP(Inter a trypsin inhibitor Heavy chain Related Protein)was found from human blood plasma by our group in 1996.About 70% of IHRP has homology in amino acid sequence to ITT heavy chain while IHRP does not show the trypsin-inhibiting activity which ITT shows. As functions of THRP has been elucidated yet, we also tried to produce IHRP gene knockout mice. Presently we have determined the structure of mouse IHRP gene as well as the nucleotide sequence of mouse IHRP mRNA.3) Studies on GBP 28While we were working with PHBP and IHRP, we also found GBP 28 from human blood plasma. GBP 28 (gelatin binding protein of 28 kd) was the translated product of apM1 mRNA which was found as an adipocyte specific mRNA by Matsuzawa et al (Osaka univ.). Matsuzawa named this protein adiponectin later. In 2001, GBP 28/adiponectin was found to have a possibility for an anti diabetes drug. Presently we are also trying to produce IHRP-gene-knockout mice.