benzyl N-[(2S)-1-[[(2S)-5-(diaminomethylideneamino)-1-[(4-methyl-2-oxochromen-7-yl)amino]-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]carbamate:hydrochloride

Description

Z-Phe-Arg-AMC (70382-26-2) is a fluorogenic substrate1 for the following cathepsins (kcat/Km in M-1s-1): B (105)2, C/DPP-I (104)3, F (106)4, K/O2 (105)3, L (106)3, L2/V (105)5, O6, S (104)7, X/Z (104)8. The peptide is also cleaved by plasma kallikrein and kallikrein 89, and papain (kcat/Km=105 M-1s-1).3Excitation: 365nm, Emission: 440nm.5

Uses

Z-Phe-Arg 7-amido-4-methylcoumarin hydrochloride has been used:

• as a fluorogenic substrate in actinidin inhibition assay
• as a kallikrein substrate
• as a trypsin substrate for fluorometric assay
• as a cathepsin-L substrate

General Description

Substrate for fluorogenic assay of plasma and glandular kallikreins. Also serves as a substrate for cathepsin B, cathepsin L, and papain.

Biochem/physiol Actions

Z-Phe-Arg 7-amido-4-methylcoumarin (Z-FR-AMC) proteolytic lysis by proteases leads to the liberation of AMC resulting in increased fluorescence in the enzymatic reaction.

References

[1] FRANCIS X. TAVARES. Design of Potent, Selective, and Orally Bioavailable Inhibitors of Cysteine Protease Cathepsin K[J]. Journal of Medicinal Chemistry, 2003, 47 3: 588-599. DOI:10.1021/jm030373l
[2] CHRISTIAN THERRIEN. Cathepsins X and B Can Be Differentiated through Their Respective Mono- and Dipeptidyl Carboxypeptidase Activities†[J]. Biochemistry Biochemistry, 2001, 40 9: 2702-2711. DOI:10.1021/bi002460a
[3] DORIT K. NÄGLER. Interdependency of Sequence and Positional Specificities for Cysteine Proteases of the Papain Family‡[J]. Biochemistry Biochemistry, 1999, 38 15: 4868-4874. DOI:10.1021/bi982632s
[4] B WANG. Human cathepsin F. Molecular cloning, functional expression, tissue localization, and enzymatic characterization.[J]. The Journal of Biological Chemistry, 1998, 273 48: 32000-32008. DOI:10.1074/jbc.273.48.32000
[5] DIETER BRÖMME. Human Cathepsin V Functional Expression, Tissue Distribution, Electrostatic Surface Potential, Enzymatic Characterization, and Chromosomal Localization‡[J]. Biochemistry Biochemistry, 1999, 38 8: 2377-2385. DOI:10.1021/bi982175f
[6] G. VELASCO. Human cathepsin O. Molecular cloning from a breast carcinoma, production of the active enzyme in Escherichia coli, and expression analysis in human tissues.[J]. The Journal of Biological Chemistry, 1994, 47 1: 27136-27142. DOI:10.1016/s0021-9258(18)47135-9
[7] G KOPITAR. Folding and activation of human procathepsin S from inclusion bodies produced in Escherichia coli.[J]. European journal of biochemistry, 1996, 236 2: 558-562. DOI:10.1111/j.1432-1033.1996.00558.x
[8] IVICA KLEMENČIČ. Biochemical characterization of human cathepsin X revealed that the enzyme is an exopeptidase, acting as carboxymonopeptidase or carboxydipeptidase[J]. The FEBS journal, 2003, 267 17: 5404-5412. DOI:10.1046/j.1432-1327.2000.01592.x
[9] TADAAKI KISHI. Activation and enzymatic characterization of recombinant human kallikrein 8.[J]. Biological Chemistry, 2006, 387 6: 723-731. DOI:10.1515/bc.2006.091