Hydrolysis of somatostatin by human tissue kallikrein after the amino acid pair phe-Phe.
Somatostatin-(1-14) was hydrolysed by human tissue kallikrein at the Phe7-Trp8 bond, after a Phe-Phe pair of amino acids, with similar kinetic parameters to those described for human high- and low-molecular-mass kininogens. Substance P and human insulin, which also contain a Phe-Phe pair in their sequences, were both resistant. More details of this hydrolytic specificity of human tissue kallikrein were obtained by synthesizing and assaying internally quenched fluorescent peptides containing the sequence of somatostatin-(1-14), as well as the reactive-centre loop of human kallikrein-binding protein (kallistatin). We also observed that human tissue kallikrein hydrolysed growth hormone-releasing hormone at the Arg11-Lys12 bond, although this peptide contains in its structure a pair of leucines (Leu22-Leu23), in contrast with the Phe-Phe pair in somatostatin. We have also demonstrated the susceptibility to human tissue kallikrein of some chromogenic peptide s with the general structure of X-Phe-Phe-p-nitroanilide and D-Pro-Phe-Phe-4-methylcoumaryl-7-amide.
Pimenta
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DC
Department of Biophysics, Escola Paulista de Medicina, Rua Três de Maio, 100 São Paulo - SP - 04044-020, Brazil.
Juliano
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MA
Juliano
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L
D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003374:Coumarins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D013007:Growth Hormone-Releasing Hormone / Q000032:analysis / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007328:Insulin / Q000378:metabolism; D007610:Kallikreins / Q000032:analysis / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis / Q000378:metabolism; D015843:Serpins / Q000737:chemistry; D013004:Somatostatin / Q000378:metabolism*; D013373:Substance P / Q000378:metabolism; D013379:Substrate Specificity; D020840:Tissue Kallikreins
D002352:Carrier Proteins; D003374:Coumarins; D005456:Fluorescent Dyes; D007328:Insulin; D010455:Peptides; D015843:Serpins; C051324:kallistatin; D013373:Substance P; D013004:Somatostatin; D013007:Growth Hormone-Releasing Hormone; D007610:Kallikreins; D020840:Tissue Kallikreins