Chromogenic Peptide Substrates
Published 1 January 1999
Heidrun Kirschke, Bernd Wiederanders
Citations74
Generate an AI Snapshot to get a quick, structured summary of this paper.
Study Snapshot
ObjectiveStudy objective
MethodsResearch methodology
PopulationPopulation studied
Sample sizeSample sizes
OutcomesStudy outcomes here
ResultsStudy results comes here
LimitationsResearch study limitations comes here
A concise AI-generated summary of the paper will appear here once you click Generate AI Snapshot.
TL;DR
This chapter deals with spectrophotometric methods for the determination of proteolytic enzymes by their chromogenic substrates and the choice of sensitive substrates for several enzymes.
Abstract
This chapter deals with spectrophotometric methods for the determination of proteolytic enzymes by their chromogenic substrates. The choice of sensitive substrates for several enzymes is facilitated by a summary of kinetic constants included in additional tables.
Keywords
Biochemistry, Genetics and Molecular Biology
Archives of Biochemistry and BiophysicsTissue sulfhydryl groups
26,215 Citations1959George L. Ellman
A water-soluble (at pH 8) aromatic disulfide [5,5′-dithiobis(2-nitrobenzoic acid] has been synthesized and shown to be useful for determination of sulfhydryl groups.
Archives of Biochemistry and BiophysicsThe preparation and properties of two new chromogenic substrates of trypsin
3,374 Citations1961Bernard F. Erlanger, Nicholas Kokowsky +1 more
Preliminary studies indicate that benzoyl dl -arginine p -nitroanilide hydrochloride is also hydrolyzed by papain, and that of l -LPA is in a more alkaline region than normally found for trypsin substrates.
Methods in enzymology on CD-ROM/Methods in enzymology[41] Cathepsin B, cathepsin H, and cathepsin L
1,692 Citations1981A. John Barrett, Heidrun Kirschke
Two types of purification methods for Cathepsin B, CathePSin H, and Cathepsypsin L are described: method I is applicable to large amounts of frozen tissues, whereas method II is used with flesh tissue and takes advantage of a 50-fold purification factor attainable by isolation of lysosomes.
Archives of Biochemistry and BiophysicsDetermination of sulfhydryl groups with 2,2′- or 4,4′-dithiodipyridine
993 Citations1967D.R. Grassetti, John F. Murray
The proposed method is compared with known methods for the determination of sulfhydryl groups and permits determination of SH groups in simple compounds and in biological materials.
Methods in enzymology on CD-ROM/Methods in enzymology[28] Assay of coagulation proteases using peptide chromogenic and fluorogenic substrates
306 Citations1981Richard Lottenberg, Ulla Christensen +2 more
The recognition that both the selectivity of many proteases and their catalytic efficiency depend on interactions with subsite amino acids in the peptide substrate coupled with the availability of amino acid sequences around the cleavage sites in several zymogens of the coagulation and fibrinolytic systems has led to the synthesis and commercial availability of a variety of peptide chromogenic and fluorogenic substrates with much greater selectivity.
BiochemistryHuman and murine cytotoxic T lymphocyte serine proteases: subsite mapping with peptide thioester substrates and inhibition of enzyme activity and cytolysis by isocoumarins
248 Citations1991Shinjiro Odake, Chih Min Kam +6 more
Although the general serine protease inhibitor 3,4-dichloroisocoumarin (DCI) inactivated these tryptases poorly, it was the best isocou marin inhibitor for murine granzyme B (kobs/[I] = 3700-4200 M-1 s-1).
Journal of Biological ChemistryNew Observations on the Substrate Specificity of Cathepsin C (Dipeptidyl Aminopeptidase I)
225 Citations1969J. Ken McDonald, Benjamin B. Zeitman +2 more
Highly purified cathepsin C was characterized as a Cl--activated dipeptidyl aminopeptidase of broader specificity than hitherto known for the bovine spleen enzyme, and substrate specificity characteristics comparable to its hydrolytic activity on amides, β-naphthylamides, and esters.
Journal of Biological ChemistrySensitive, soluble chromogenic substrates for HIV-1 proteinase.
199 Citations1990Anthony D. Richards, Lowri H. Phylip +8 more
The spectrophotometric assay has been used to characterize the proteinase further with respect to pH dependence, ionic strength dependence, and the effect of competitive inhibitors of various types.
Biochemical JournalA systematic series of synthetic chromophoric substrates for aspartic proteinases
176 Citations1986Ben M. Dunn, Mar x ED a de la Paz Jimenez +4 more
The Phe-Phe(4-NO2) bond was established as the only peptide bond cleaved, and kinetic constants were obtained for the hydrolysis of these peptide substrates by a representative selection of aspartic proteinases of animal and microbial origin.
BiochemistrySubstrate specificity and kinetic characteristics of angiotensin converting enzyme
116 Citations1983Peter Buenning, Barton Holmquist +1 more
The substrate specificity of the enzyme has been examined in terms of the fundamental specificity quantity kcat/KM as well as the separate constants by using a series of furanacryloyl-tripeptides and has been compared with that toward the physiological substrates angiotensin I and bradykinin.
Journal of Biological ChemistryMammalian tissue trypsin-like enzymes. Comparative reactivities of human skin tryptase, human lung tryptase, and bovine trypsin with peptide 4-nitroanilide and thioester substrates.
102 Citations1983Takumi Tanaka, Brian J. McRae +5 more
The preference of the skin tryptase for substrates with two terminal basic residues indicates that this enzyme could process prohormones and proproteins which contain this structural feature at the cleavage site.
Methods in enzymology on CD-ROM/Methods in enzymology[42] Crystalline leucine aminopeptidase from lens (α-aminoacyl-peptide hydrolase; EC 3.4.11.1)
101 Citations1976Horst Hanson, M Frohne
Biochemical JournalHuman cathepsin B. Application of the substrate <i>N</i>-benzyloxycarbonyl-<scp>l</scp>-arginyl-<scp>l</scp>-arginine 2-naphthylamide to a study of the inhibition by leupeptin
90 Citations1980Christopher G. Knight
Journal of Histochemistry & CytochemistryLysosomal cysteine and aspartic proteinases, acid phosphatase, and an endogenous cysteine proteinase inhibitor, cystatin-beta, in rat osteoclasts.
82 Citations1993Yoshiyuki Ohsawa, Tohru Nitatori +3 more
The results suggest that lysosomal cysteine and aspartic proteinases may play a role in the degradation of organic constituents of the bone matrix and cystatin-beta can serve as an excellent marker protein for osteoclasts.
Biochemical JournalCathepsin S The cysteine proteinase from bovine lymphoid tissue is distinct from cathepsin L (EC 3.4.22.15)
74 Citations1986Heidrun Kirschke, Isabelle Schmidt +1 more
Preparations of cathepsin L from rat liver, rat kidney and bovine liver were shown to have kinetic constants for the substrate benzyloxycarbonyl-Phe-Arg-7-(4-methyl)coumarylamide in the same range (Km 2-3 microM).
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular EnzymologyPhotometric or fluorometric assay of cathepsin B, L and H and papain using substrates with an aminotrifluoromethylcoumarin leaving group
72 Citations1991Jean Roger Tchoupé, Thierry Moreau +2 more
Benzyloxycarbonyl-Phe-Arg-N-trifluoromethylcoumarinylamide is a highly sensitive substrate for papain and can be used as a chromogenic substrate for cathepsin B, L and H.
Methods in enzymology on CD-ROM/Methods in enzymology[18] Carboxypeptidases C and D
52 Citations1994S. James Remington, Klaus Breddam
This chapter examines carboxypeptidases C and D, members of serine carboxypesptidase family that specifically release amino acids from the C termini of peptides and proteins.
FEBS LettersSelective cleavage of glycyl bonds by papaya proteinase IV
48 Citations1990David J. Buttle, Anka Ritonja +3 more
The structure of PPIV has been modelled on that of papain, and it is suggested that the replacement of the highly conserved residues Gly‐65 and Gly‐23 by arginine and glutamic acid can account for the specificity ofPPIV.
BiochemistrySite-directed mutagenesis on (serine) carboxypeptidase Y. A hydrogen bond network stabilizes the transition state by interaction with the C-terminal carboxylate group of the substrate
41 Citations1994Uffe Hasbro Mortensen, S. James Remington +1 more
The three-dimensional structure of (serine) carboxypeptidase Y suggests that the side chains of Trp49, Asn51, Glu65, and Glu145 could be involved in the recognition of the C-terminal carboxylate group of peptide substrates, but strong interactions are formed only in the transition state.
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular EnzymologyElastolytic activity of Pseudomonas aeruginosa elastase
37 Citations1989Joëlle Saulnier, Florence M. Curtil +2 more
P. aeruginosa elastase may hydrolyse tetrapeptides such as tetraalanine, or synthetic substrates such as furylacryloyltripeptides FA-X-Leu-Y, X and Y being Gly and/or Ala.
Journal of Biological ChemistryCathepsins B and H from porcine spleen. Purification, polypeptide chain arrangements, and carbohydrate content.
34 Citations1984Takayuki Takahashi, A H Dehdarani +2 more
Procedures for the purification of cathepsins B and H from porcine spleens have been described and showed that these enzymes were glycoproteins.
Biochemical JournalEnzyme-substrate interactions in the hydrolysis of peptides by cathepsins B and H from rat liver
32 Citations1987Dieter Brömme, K Bescherer +2 more
The number of possible subsites of the rat liver cysteine proteinases cathepsin B and cat hepsin H was determined in the N-terminal direction from the scissile bond to show a preference for the amide substrate and an elongation of the substrate peptide chain of up to four amino acid residues enhances the hydrolysis rate.
Methods in enzymology on CD-ROM/Methods in enzymology[1] Peptide thioester substrates for serine peptidases and metalloendopeptidases
27 Citations1995James C. Powers, Chih-Min Kam
Tripeptide substrates of the type Boc-Ala-AlA-AA-SBzl, where AA is nearly all of the 20 common amino acids, have now been synthesized and should be useful for the subsite mapping of new serine peptidases and the study of crude cell preparations containing serinepeptidases.
Methods in enzymology on CD-ROM/Methods in enzymology[16] Dipeptidyl-peptidase IV from rat liver
23 Citations1994Yukio Ikehara, Shigenori Ogata +1 more
This chapter describes assay, purification, and properties of diPEptidyl-peptidase IV (DPPIV) from rat liver, a serine protease that cleaves N-terminal dipeptides from oligo- and polypeptide with a penultimate prolyl residue.
Biochemical JournalBenzyloxycarbonylphenylalanylcitrulline <i>p</i>-nitroanilide as a substrate for papain and other plant cysteine proteinases
22 Citations1984C.J. Gray, John Boukouvalas +2 more
It is concluded that benzyloxycarbonylphenylalanylcitrulline p-nitroanilide is a highly selective substrate for the sensitive detection and assay of the plant cysteine proteinases.
Clinical Chemistry and Laboratory Medicine (CCLM)Isolation of Human Urinary Kallikrein by Affinity Chromatography. Determination of human urinary kallikrein, I.
14 Citations1977R. Geiger, K. Mann +1 more
The kinetic constants for hydrolysis of BZArgOEt, TosArgOMe, CbzTyrONp, and human HMW-kininogen were determined and Benzamidine and aprotinin are competitive inhibitors of human urinary kallikrein.
Archives of Biochemistry and BiophysicsThioester Hydrolysis by Matrix Metalloproteinases
13 Citations1994Ross L. Stein, Maria Izquierdo-Martin
The pH-dependence of kc/Km for the SLN-catalyzed hydrolysis of Ac-Pro-Leu-Ala-SNva-TrpNH2 is identical to pH-dependencies for peptide Hydrolysis and suggests no major mechanistic differences between thioester and amide hydrolyzing by SLN.
Clinical Chemistry and Laboratory Medicine (CCLM)Photometrische Bestimmung der „klassischen“ Leucinaminopeptidase im Blutplasma und -serum
13 Citations1968Reinhard J. Haschen, W Farr +1 more
Hier zeigt sich erneut, daß bei der Interpretation biologischer Erscheinungen die statistische Signifikanz sehr wohl von der Relevanz des beobachteten Effektes unterschieden werden muß wird die Rolle der Niere zu berücksichtigen sein.
The Journal of BiochemistryPurification and Characterization of a Novel Acrosin-Like Enzyme from Boar Cauda Epididymal Sperm
11 Citations1994Kuniko Akama, Kimio Terao +5 more
The results suggest that CSP is a novel acrosin-like enzyme that differs from acrosIn in its location in the sperm head, the effect of calcium ions on its activity, and its substrate specificity.