tailieunhanh - Báo cáo khóa học: Development of recombinant inhibitors specific to human kallikrein 2 using phage-display selected substrates

The reactive site loop of serpins undoubtedly defines in part their ability to inhibit a particular enzyme. Exchanges in the reactive loop of serpins might reassign the targets and modify the serpin–protease interaction kinetics. Based on this concept, we have developed a procedure to change the specificity of known serpins. First, reactive loops are very good substrates for the target enzymes. Therefore, we have used the phage-display technology to select from a penta-peptide phage library the best substrates for the human prostatekallikreinhK2 . | Eur. J. Biochem. 271 607-613 2004 FEBS 2004 doi Development of recombinant inhibitors specific to human kallikrein 2 using phage-display selected substrates Sylvain M. Cloutier1 2 Christoph Kiindig2 Loyse M. Felber1 Omar M. Fattah1 Jair R. Chagas3 Christian M. Gygi1 Patrice Jichlinski1 Hans-Jurg Leisinger1 and David Deperthes1 2 1 Urology Research Unit Department of Urology CHUV Epalinges Switzerland 2Med Discovery SA Epalinges Switzerland 3Centro Interdisciplinar de Investigacao Bioquimica Universidade de Mogi das Cruzes Brazil The reactive site loop of serpins undoubtedly defines in part their ability to inhibit a particular enzyme. Exchanges in the reactive loop of serpins might reassign the targets and modify the serpin-protease interaction kinetics. Based on this concept we have developed a procedure to change the specificity of known serpins. First reactive loops are very good substrates for the target enzymes. Therefore we have used the phage-display technology to select from a pentapeptide phage library the best substrates for the human prostate kallikrein hK2 Cloutier . Chagas . Mach . Gygi . Leisinger . Deperthes D. 2002 Eur. J. Biochem. 269 2747-2754 . Selected substrates were then transplanted into the reactive site loop of a1-antichymotrypsin to generate new variants of this serpin able to inhibit the serine protease. Thus we have developed some highly specific a1-antichymotrypsin variants toward human kallikrein 2 which also show high reactivity. These inhibitors might be useful to help elucidate the importance of hK2 in prostate cancer progression. Keywords phage-display protease human kallikrein inhibitor a1-antichymotrypsin. Prostate cancer is currently the most commonly diagnosed cancer in American men. This pathology is the second leading cause of cancer death after lung cancer and the majority of the patients with locally advanced prostate cancer have an increased risk for disease progression. In .