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Literature DB >> 3593241

An improved spectrophotometric assay for leucine aminopeptidase.

R E Beattie, D J Guthrie, D T Elmore, C H Williams, B Walker.

Abstract

A sensitive assay to determine the activity of leucine aminopeptidase (EC 3.4.11.1), using L-leucine thiobenzyl ester as substrate, was developed. Hydrolysis of the ester by leucine aminopeptidase can be monitored in the presence of 5,5-dithiobis-(2-nitrobenzoic acid) by continuous spectrophotometric measurement at 412 nm. Comparison with some amide substrates showed that the thiol ester provides a much more sensitive assay, its specificity constant (Vmax./Km) being some 3000-fold higher than that of leucine p-nitroanilide.

Entities: Chemical

Mesh：

Substances：

Year: 1987 PMID： 3593241 PMCID： PMC1147694 DOI： 10.1042/bj2420281

Source DB: PubMed Journal: Biochem J ISSN： 0264-6021 Impact factor: 3.857

20 in total

1. Spectrophotometric assay of peptidase activity.

Authors: F BINKLEY; C TORRES
Journal: Arch Biochem Biophys Date: 1960-02 Impact factor: 4.013

2. Crystalline leucine aminopeptidase from lens (alpha-aminoacyl-peptide hydrolase; EC 3.4.11.1).

Authors: H Henson; M Frohne
Journal: Methods Enzymol Date: 1976 Impact factor: 1.600

3. A new fluorogenic substrate for chymotrypsin.

Authors: M Zimmerman; E Yurewicz; G Patel
Journal: Anal Biochem Date: 1976-01 Impact factor: 3.365

4. Use of N-benzoyl-L-tyrosine thiobenzyl ester as a protease substrate. Hydrolysis by alpha-chymotrypsin and subtilisin BPN.

Authors: D A Farmer; J H Hageman
Journal: J Biol Chem Date: 1975-09-25 Impact factor: 5.157

5. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples.

Authors: M A Markwell; S M Haas; L L Bieber; N E Tolbert
Journal: Anal Biochem Date: 1978-06-15 Impact factor: 3.365

6. Leucine aminopeptidase from swine kidney: purification, molecular weight, subunit and amino acid composition.

Authors: C Shen; P Mellius
Journal: Prep Biochem Date: 1977

7. Thiobenzyl benzyloxycarbonyl-L-lysinate, substrate for a sensitive colorimetric assay for trypsin-like enzymes.

Authors: G D Green; E Shaw
Journal: Anal Biochem Date: 1979-03 Impact factor: 3.365

8. Sensitive substrates for human leukocyte and porcine pancreatic elastase: a study of the merits of various chromophoric and fluorogenic leaving groups in assays for serine proteases.

Authors: M J Castillo; K Nakajima; M Zimmerman; J C Powers
Journal: Anal Biochem Date: 1979-10-15 Impact factor: 3.365