scholarly journals Quantification of a matrix metalloproteinase-generated aggrecan G1 fragment using monospecific anti-peptide serum

1995 ◽  
Vol 307 (1) ◽  
pp. 245-252 ◽  
Author(s):  
M W Lark ◽  
H Williams ◽  
L A Hoernner ◽  
J Weidner ◽  
J M Ayala ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Matrix Metalloproteinase ◽  
Limit Of Detection ◽  
Cartilage Destruction ◽  
Cathepsin G ◽  
Terminal Sequence ◽  
Leucocyte Elastase ◽  
The Matrix ◽  
Radio Immunoassay ◽  
Species Specific

Several members of the matrix metalloproteinase family have been reported to cleave aggrecan in the interglobular domain between Asn-341 and Phe-342. An antiserum was prepared against a peptide conjugate corresponding to the C-terminal sequence of the matrix metalloproteinase-generated aggrecan G1 fragment (Phe335-Val-Asp-Ile-Pro-Glu-Asn341). A quantitative radioimmunoassay, with a limit of detection of about 80 pM, was developed using this antiserum. This antiserum requires the free carboxyl group of the C-terminal asparagine for optimal recognition. If the C-terminal asparagine is excised from the sequence, replaced with closely related amino acids, or extended across the matrix metalloproteinase cleavage site, there is a 40-10,000-fold loss in detection. Using peptides cleaved from the N-terminus, it was determined that the antiserum requires the entire Phe-Val-Asp-Ile-Pro-Glu-Asn sequence for optimal recognition. The radioimmunoassay detects matrix metalloproteinase-generated G1 fragments with similar sensitivity to the Phe-Val-Asp-Ile-Pro-Glu-Asn peptide, but it does not recognize intact aggrecan. Immunoreactive aggrecan G1 fragments of molecular mass 50 kDa are generated by the matrix metalloproteinases stromelysin and gelatinase A. In contrast, under identical conditions, the closely related metalloproteinases, gelatinase B and collagenase, as well as cathepsin G, cathepsin B and human leucocyte elastase, did not generate a G1 fragment recognized by the antiserum. The anti-Phe-Val-Asp-Ile-Pro-Glu-Asn serum detects stromelysin-generated aggrecan G1 fragments from mouse, guinea pig, rabbit and human, indicating that the detection is not species-specific. This antiserum and radio-immunoassay should be useful for quantifying and characterizing matrix metalloproteinase-generated aggrecan G1 fragments in articular cartilage and synovial fluids from humans and various animal models of articular-cartilage destruction.

scholarly journals Purification of a serine-proteinase inhibitor from human articular cartilage. Identity with the acid-stable proteinase inhibitor of mucous secretions

1991 ◽  
Vol 274 (1) ◽  
pp. 269-273 ◽  
Author(s):  
B Böhm ◽  
R Deutzmann ◽  
H Burkhardt
Keyword(s):  
Articular Cartilage ◽  
Proteinase Inhibitor ◽  
Serine Proteinase ◽  
Cathepsin G ◽  
Human Articular Cartilage ◽  
Serine Proteinase Inhibitor ◽  
Inhibitor Molecule ◽  
Leucocyte Elastase ◽  
Sds Page

An inhibitor of the serine proteinases human leucocyte elastase (EC 3.4.21.37), of cathepsin G (EC 3.4.21.20) and of trypsin (EC 3.4.21.4) has been purified from human articular cartilage. The apparent Mr of the cationic (pI greater than 10) protein was determined to 15,000 by SDS/PAGE. It was shown to cross-react in Western blot with a specific antibody to a recombinant-derived serine-proteinase inhibitor of human mucous secretions. Identity of both inhibitors is indicated by the determination of the N-terminal amino acid sequence of the cartilage-derived serine-proteinase inhibitor. In all 24 residues the cartilage inhibitor was shown to be identical with the human secretory leucocyte proteinase inhibitor (‘SLPI’). The inhibitor molecule may play a crucial role in the protection of cartilage matrix proteins against proteolytic attack.

The effect of thymosin β4 on articular cartilage chondrocyte matrix metalloproteinase expression

2002 ◽  
Vol 30 (6) ◽  
pp. 879-882 ◽  
Author(s):  
E. J. Blain ◽  
D. J. Mason ◽  
V. C. Duance
Keyword(s):  
Articular Cartilage ◽  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Mechanical Loading ◽  
Functional Role ◽  
Fold Increase ◽  
Thymosin Β4 ◽  
Cytoskeletal Organization ◽  
Primary Chondrocytes ◽  
The Matrix

Mechanical loading is paramount in regulating both the anabolic and catabolic activities of articular cartilage chondrocytes, essential for the matrix to retain its functional integrity. We have identified thymosin β4 as a putative mechanically regulated gene that may mediate load-enhanced synthesis and activation of matrix metalloproteinases (MMPs) 2 and 9 in articular cartilage. The objective of this study was to confirm the mechanical regulation of thymosin β4 and determine its effect on cartilage chondrocyte MMP production. Thymosin β4 mRNA expression, analysed by quantitative PCR, revealed a significant 20-fold increase in cartilage loaded for 10 min which was still evident after 30 min of loading. Treatment of primary chondrocytes with 2 and 4 μg · ml-1 thymosin β4 peptide for 4 h significantly increased pro-MMP 9 expression and activation. We postulate a functional role for load-induced thymosin β4 in modulating the cytoskeletal organization of articular cartilage chondrocytes to affect MMP expression.

scholarly journals Structure and expression of the human gene for the matrix metalloproteinase matrilysin.

1994 ◽  
Vol 269 (3) ◽  
pp. 2032-2040
Author(s):  
M. Gaire ◽  
Z. Magbanua ◽  
S. McDonnell ◽  
L. McNeil ◽  
D.H. Lovett ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Human Gene ◽  
The Matrix

scholarly journals Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1577
Author(s):  
Klaudia Kotecka-Majchrzak ◽  
Natalia Kasałka-Czarna ◽  
Agata Sumara ◽  
Emilia Fornal ◽  
Magdalena Montowska
Keyword(s):  
Limit Of Detection ◽  
Raw Materials ◽  
Meat Products ◽  
Food Products ◽  
Plant Raw Materials ◽  
Heat Stable ◽  
2S Albumins ◽  
Specific Proteins ◽  
Meat Species ◽  
Species Specific

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.

Identification for adulteration of beef with chicken based on single primer-triggered isothermal amplification

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Jiao Chen ◽  
Pansong Zhang ◽  
Haixia Wang ◽  
Yanjing Shi
Keyword(s):  
Limit Of Detection ◽  
Chicken Meat ◽  
Isothermal Amplification ◽  
Work Flow ◽  
Acid Extraction ◽  
Specificity And Sensitivity ◽  
Total Dna ◽  
Meat Species ◽  
Species Specific ◽  
Single Primer

Abstract Adulteration of beef with cheap chicken has become a growing problem worldwide. In this study, a quick, single primer-triggered isothermal amplification (SAMP) combined with a fast nucleic acid extraction method was employed to detect the chicken meat in adulterated beef. Chicken from adulterated beef was identified using the chicken species-specific primer designed according to the Gallus gallus mitochondrial conserved sequences. Our SAMP method displayed good specificity and sensitivity in detecting chicken and beef meat DNA–the limit of detection (LOD) of SAMP is 0.33 pg/μL of chicken and beef total DNA and 2% w/w chicken meat in beef. The whole work flow from DNA extraction to signal detection can be finished within 1 h, fulfilling the requirement of on-site meat species identification.

Fluorescence imaging to assess the matrix metalloproteinase activity and its inhibitor in vivo

2008 ◽  
Vol 13 (1) ◽  
pp. 011006 ◽  
Author(s):  
Zhihong Zhang ◽  
Jie Yang ◽  
Jinling Lu ◽  
Juqiang Lin ◽  
Shaoqun Zeng ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Fluorescence Imaging ◽  
The Matrix ◽  
Metalloproteinase Activity
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