caa a22 4500 475743830 CHVBK 20180406123507.0 cr unu---uuuuu 170329e20000101xx s 000 0 eng 10.1023/A:1007090708945 doi (NATIONALLICENCE)springer-10.1023/A:1007090708945 Specificity of Cathepsin B to Fluorescent Substrates Containing Benzyl Side-Chain-Substituted Amino Acids at P1 Subsite [Elektronische Daten] [Elaine Del Nery, Lira Alves, Robson Melo, Maria Cesari, Luiz Juliano, Maria Juliano] We have determined the kinetic parameters for the hydrolysis by cathepsin B of peptidyl-coumarin amide and intramolecularly quenched fluorogenic peptides with the general structures εNH2-Cap-Leu-X-MCA and Abz-Lys-Leu-X-Phe-Ser-Lys-Gln-EDDnp, respectively. Abz (ortho-aminobenzoic acid) and EDDnp (2,4-dinitrophenyl-ethylenediamine) are the fluorescent donor-acceptor pair, and X was Cys(SBzl), Ser(OBzl), and Thr(OBzl) containing benzyl group (Bzl) at the functional side chain of Cys, Ser, and Thr. The peptidyl-coumarin-containing Cys(SBzl), Ser(OBzl), and Thr(OBzl) have higher affinity cathepsin B, supporting the interpretation of the crystal structure of rat cathepsin B complexed with the inhibitor Z-Arg-Ser(OBzl)-CH2Cl that the benzyl group attached to Ser hydroxyl side chain occupies the enzyme S′1 subsite [Jia et al. (1995), J. Biol. Chem. 270, 5527]. A similar effect of benzyl group was also detected in the internally quenched peptides. Finally, the benzyl group in substrates containing Cys(SBzl) amino acid at P1 seems to compensate the absence of adequate S2-P2 interaction in the hydrolysis of the peptides having Pro or Ala at P2 position. Plenum Publishing Corporation, 2000 Thiolprotease nationallicence cathepsins B nationallicence papain nationallicence fluorogenic substrate nationallicence fluorescent peptides nationallicence proteinases nationallicence Del Nery Elaine Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut Alves Lira Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut Melo Robson Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut Cesari Maria Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut Juliano Luiz Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut Juliano Maria Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/1(2000-01-01), 33-38 0277-8033 19:1<33 2000 19 10930 https://doi.org/10.1023/A:1007090708945 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007090708945 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Del Nery Elaine Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut NATIONALLICENCE 700 1- Alves Lira Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut NATIONALLICENCE 700 1- Melo Robson Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut NATIONALLICENCE 700 1- Cesari Maria Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut NATIONALLICENCE 700 1- Juliano Luiz Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut NATIONALLICENCE 700 1- Juliano Maria Department of Biophysics, Escola Paulista de Medicina, UNIFESP, 04044-020, São Paulo, Brazil aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/1(2000-01-01), 33-38 0277-8033 19:1<33 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer