| Highly specific substrates of proteinase 3 containing 3-(2-benzoxazol-5-yl)-l-alanine and their application for detection of this enzyme in human serum. | |
| | |
MedLine Citation:
|
PMID: 20361767 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
A set of benzoxazolyl-l-alanine derivates along with the MCA moiety (donors of fluorescence) were introduced into a proteinase 3 (PR3) substrate with a C-terminal ANB-NH(2) that serves as a fluorescence acceptor. Five substrates with general formula X-Tyr-Tyr-Abu-ANB-NH(2) were synthesized, and their kinetic parameters against proteinase 3 were determined. The highest k(cat)/K(M) value, 1.5 x 10(6) M(-1) s(-1), was obtained for (Pyr)Box-Ala-Tyr-Tyr-Abu-NH(2) where (Pyr)Box-Ala stands for N-methylpyrrole benzoxazole-l-alanine. Titration of this peptide with proteinase 3 resulted in measurable fluorescence at an enzyme amount equal to 29 pmol. This substrate was selected used to detect quantifiable levels of proteinase 3 in serum samples, including those of normal subjects. For all c-ANCA-positive samples (diagnosed Wegener granulomatosis), a significant increase of PR3 concentration was observed. Wegener granulomatosis is a severe autoimmune disease causing inflammation of the blood vessels. Our results clearly show that this substrate can be used for the construction of a very reliable, inexpensive, and easy to use diagnostic test for PR3 determination. |
| | |
Authors:
|
Magdalena Wysocka; Adam Lesner; Katarzyna Guzow; Julia Kulczycka; Anna Łegowska; Wiesław Wiczk; Krzysztof Rolka |
Related Documents
:
|
20116077 - Biochemical comparison of two proteolytic enzymes from carica candamarcensis: structura... 11368647 - Cathepsin l: a predominant heat-activated proteinase in arrowtooth flounder muscle. 9538257 - Purification and characterization of myonase from x-chromosome linked muscular dystroph... 6397997 - L-pyroglutamyl-l-phenylalanyl-l-leucine-p-nitroanilide--a chromogenic substrate for thi... 8556737 - Metabolism of xyloglucan generates xylose-deficient oligosaccharide subunits of this po... 7643537 - Metalloproteinase generation by human glomerular epithelial cells. |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
|
Title: Analytical chemistry Volume: 82 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2010 May |
Date Detail:
|
Created Date: 2010-04-30 Completed Date: 2010-08-18 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
|
Languages: eng Pagination: 3883-9 Citation Subset: IM |
Affiliation:
|
Faculty of Chemistry, University of Gdanìsk, Poland. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Alanine
/
chemistry* Benzoxazoles / chemistry* Blood Chemical Analysis / methods* Diagnostic Techniques and Procedures / economics, standards Humans Molecular Structure Myeloblastin / blood* Reproducibility of Results Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Substrate Specificity |
| Chemical | |
Reg. No./Substance:
|
0/Benzoxazoles; 56-41-7/Alanine; EC 3.4.21.76/Myeloblastin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Urea-Induced Drying of Carbon Nanotubes Suggests Existence of a Dry Globule-like Transient State Dur...
Next Document: Studies of Single Aerosol Particles Containing Malonic Acid, Glutaric Acid, and Their Mixtures with ...