Kurschat P, Graeve L, Erren A, Gatsios P, Rose-John S, Roeb E, Tschesche H, Koj A, Heinrich P C
Institut für Biochemie der RWTH Aachen, Germany.
Eur J Biochem. 1995 Dec 1;234(2):485-91. doi: 10.1111/j.1432-1033.1995.485_b.x.
Murine tissue inhibitor of metalloproteinases-1 (mTIMP-1) was expressed in baculovirus-infected insect cells (Sf9). The protein secreted into the culture medium was purified to homogeneity by means of heparin-Sepharose CL-6B and FPLC. The purified protein showed metalloproteinase-inhibitory activity in two independent assays: reverse zymography and inhibition of collagenase activity. Digestion of the recombinant TIMP-1 with peptide-N-glycanaseF revealed that both N-glycosylation sites are used. 125I-mTIMP-1 intraveneously injected into a male Sprague Dawley rat disappeared within 2 min from the circulation. 5 min after injection more than 50% of the 125I-mTIMP-1 were found in the liver and 20% in the kidneys. At later times, trichloroacetic-acid-soluble material accumulated in the intestinal tract.
小鼠金属蛋白酶组织抑制剂-1(mTIMP-1)在杆状病毒感染的昆虫细胞(Sf9)中表达。分泌到培养基中的蛋白质通过肝素-琼脂糖CL-6B和快速蛋白质液相色谱法纯化至同质。纯化后的蛋白质在两项独立试验中显示出金属蛋白酶抑制活性:反向酶谱法和胶原酶活性抑制。用肽-N-聚糖酶F消化重组TIMP-1表明两个N-糖基化位点均被利用。静脉注射到雄性斯普拉格-道利大鼠体内的125I-mTIMP-1在2分钟内从循环中消失。注射后5分钟,超过50%的125I-mTIMP-1在肝脏中被发现,20%在肾脏中。在随后的时间里,三氯乙酸可溶性物质在肠道中积累。