Comparative substrate specificity analysis of recombinant human cathepsin V and cathepsin L

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• 作者：Puzer ; Luciano ; Cotrin ; Simone S. ; Alves ; Marcio F.M. ; Egborge ; Tobore ; Ara脙脙jo ; Mariana S. ; et. al.
• 关键词：Lysosomal proteases ; Cysteine proteases ; Cathepsin L ; Cathepsin V ; Fluorogenic substrates ; Bradykinin ; Kininogen
• 刊名：Archives of Biochemistry and Biophysics
• 出版年：2004
• 期刊代码：116_00039861
• 类别：ch
• 出版时间：October 15, 2004
• 卷：430
• 期：2
• 页码：274-283
• 文件大小：322 K

摘要

Cathepsins V and L have high identity and few structural differences. In this paper, we reported a comparative study of the hydrolytic activities of recombinant human cathepsins V and L using fluorescence resonance energy transfer peptides derived from Abz-KLRSSKQ-EDDnp (Abz=ortho-aminobenzoic acid and EDDnp=N-(2,4-dinitrophenyl)ethylenediamine). Five series of peptides were synthesized to map the S₃ to Formula Not Shown subsites. The cathepsin V subsites S₁ and S₃ present a broad specificity while cathepsin L has preference for positively charged residues. The S₂ subsites of both enzymes require hydrophobic residues with preference for Phe and Leu. The Formula Not Shown and Formula Not Shown subsites of cathepsins V and L are less specific. Based on these data we designed substrates to explore the electrostatic potential differences of them. Finally, the kininogenase activities of these cathepsins were compared using synthetic human kininogen fragments. Cathepsin V preferentially released Lys-bradykinin while cathepsin L released bradykinin. This kininogenase activity by cathepsins V and L was also observed from human high and low molecular weight kininogens.