scholarly journals Resistance of small leucine-rich repeat proteoglycans to proteolytic degradation during interleukin-1-stimulated cartilage catabolism

1999 ◽  
Vol 339 (3) ◽  
pp. 571-577 ◽  
Author(s):  
Robert SZTROLOVICS ◽  
Robert J. WHITE ◽  
A. Robin POOLE ◽  
John S. MORT ◽  
Peter J. ROUGHLEY
Keyword(s):  
Degradation Product ◽  
Culture Media ◽  
Degradation Products ◽  
Interleukin 1 ◽  
Culture Period ◽  
Proteolytic Degradation ◽  
Early Event ◽  
Globular Domain ◽  
Leucine Rich Repeat ◽  
Nasal Cartilage

A bovine nasal-cartilage culture system has been utilized to analyse the catabolic events occurring in response to interleukin-1β over a 14-day period. An early event following the start of interleukin-1 treatment was the release of glycosaminoglycan into the culture medium. This release was accompanied by the appearance in the tissue, and shortly thereafter also in the culture media, of a globular domain (G1)-containing aggrecan degradation product generated by the action of aggrecanase. Link protein was also released from the cartilage with a similar timeframe to that of the G1 fragment, although there was no evidence of its proteolytic degradation. By comparison with aggrecan, the small leucine-rich repeat proteoglycans decorin, biglycan and lumican showed a resistance to both proteolytic cleavage and release throughout the culture period. In contrast, fibromodulin exhibited a marked decrease in size after day 4, presumably due to proteolytic modification, but the major degradation product was retained throughout the culture period. Also in contrast with the early changes in the components of the proteoglycan aggregate, type II collagen did not display signs of extensive degradation until much later in the culture period. Collagen degradation products compatible with collagenase action first appeared in the medium by day 10 and increased thereafter. These data demonstrate that the leucine-rich repeat proteoglycans are resistant to proteolytic action during interleukin-1-stimulated cartilage catabolism, compared with aggrecan. This resistance and continued interaction with the surface of the collagen fibrils may help to stabilize the collagen fibrillar network and protect it from extensive proteolytic attack during the early phases of cartilage degeneration.

scholarly journals The degradation of cartilage proteoglycans by tissue proteinases. Proteoglycan heterogeneity and the pathway of proteolytic degradation

1977 ◽  
Vol 167 (3) ◽  
pp. 639-646 ◽  
Author(s):  
P J Roughley
Keyword(s):  
Cathepsin B ◽  
Degradation Products ◽  
Chondroitin Sulphate ◽  
Cathepsin G ◽  
Proteolytic Degradation ◽  
Single Chain ◽  
Structural Variations ◽  
Nasal Cartilage ◽  
Core Proteins ◽  
Average Size

1. CaCl2-extracted proteoglycan from bovine nasal cartilage was degraded by four tissue proteinases till no further decrease in hydroynamic size was obtained. The proteoglycan and its final degradation products were then fractionated by Sepharose 2B chromatography. 2. The average size of the degradation products was least for cathepsin B and lysosomal elastase, and greatest for cathepsin D and cathepsin G. The latter two proteinases also produced degradation products that showed the widest range of sizes. 3. The structure of the degradation products ranged from peptides containing a single glycosaminoglycan chain to those containing twelve or more chains. Of the four proteinases, only cathepsin B produced peptides that contained a single chondroitin sulphate chain. 4. The proteoglycan was very heterogeneous with respect to size and chemical composition. Its behaviour on electrophoresis suggested that at least two genetically distinct core proteins might exist. 5. Irrespective of their structural variations, all proteoglycan molecules were able to interact with hyaluronic acid. In contrast, none of the degradation products were capable of this type of interaction. 6. A pathway for the proteolytic degradation of proteoglycans is postulated in which the sites of initial cleavage may be common to the majority of proteinases, whereas the production of the final clusters is dependent on the specificity of the proteinase. Only those proteinases of broadest specificity can produce single-chain chondroitin sulphate-peptides.

scholarly journals Resistance of small leucine-rich repeat proteoglycans to proteolytic degradation during interleukin-1-stimulated cartilage catabolism

1999 ◽  
Vol 339 (3) ◽  
pp. 571 ◽  
Author(s):  
Robert SZTROLOVICS ◽  
Robert J. WHITE ◽  
A. Robin POOLE ◽  
John S. MORT ◽  
Peter J. ROUGHLEY
Keyword(s):  
Interleukin 1 ◽  
Proteolytic Degradation ◽  
Leucine Rich Repeat

scholarly journals Differences in susceptibility of marine bacterial communities to metal pyrithiones, their degradation compounds and organotin antifouling biocides

2019 ◽  
Vol 99 (5) ◽  
pp. 1033-1039
Author(s):  
Madoka Ohji ◽  
Hiroya Harino ◽  
William John Langston
Keyword(s):  
Life History ◽  
Degradation Product ◽  
Bacterial Communities ◽  
Degradation Products ◽  
High Susceptibility ◽  
Zinc Pyrithione ◽  
Bacterial Colony ◽  
Indigenous Bacteria ◽  
Antifouling Biocides ◽  
Colony Forming Units

AbstractThe susceptibility of marine bacterial communities to copper pyrithione (CuPT2), zinc pyrithione (ZnPT2) and their degradation product is described and toxicities of these relatively new antifouling biocides compared with those of their harmful organotin (OT) predecessors, tributyltin (TBT) and triphenyltin (TPT). These biocides were added to agar at concentrations of 0, 0.01, 0.1, 1 and 10 mg l−1and coastal seawater including indigenous bacteria added to each batch of agar solution. The number of bacterial colony forming units (CFU) was measured after 7 days culture. Relative CFU (as a percentage of control) was more than 80% at a concentration of 0.01 mg l−1of each compound, except for TBT. Relative CFU decreased as a function of dose of each biocide, although concentration-dependent changes in rate of CFU were relatively low during exposure to degradation products of CuPT2and ZnPT2, pyridineN-oxide (PO) and pyridine-2-sulphonic acid (PSA). Based on comparisons of EC50, TBT was the most bacterio-toxic of the tested compounds (0.2 mg l−1), marginally more so than CuPT2(0.3 mg l−1). Interestingly, EC50values of degradation products of CuPT2and ZnPT2, 2-mercaptopyridineN-oxide (HPT) and 2,2′-dithio-bispyridineN-oxide (PT2) were 0.8 and 0.5 mg l−1, respectively, lower than that of the parent chemical, ZnPT2(1.4 mg l−1). The EC50of PT2was also lower than that of TPT (0.7 mg l−1), implying higher toxicity. Given the overlapping toxicity ranges, these results suggest that marine bacterial communities experience comparably high susceptibility to metal PTs and OTs during their life history.

scholarly journals Intestinal and Hepatic Uptake of Dietary Peroxidized Lipids and Their Decomposition Products, and Their Subsequent Effects on Apolipoprotein A1 and Paraoxonase1

Antioxidants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1258
Author(s):  
Xueting Jiang ◽  
Pragney Deme ◽  
Rajat Gupta ◽  
Dmitry Litvinov ◽  
Kathryn Burge ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Degradation Products ◽  
Apolipoprotein A1 ◽  
Paraoxonase 1 ◽  
Pon1 Activity ◽  
Atherosclerotic Cardiovascular Disease ◽  
Metabolic Fate ◽  
Cell Culture Media ◽  
Decomposition Products

Both pro- and antiatherosclerotic effects have been ascribed to dietary peroxidized lipids. Confusion on the role of peroxidized lipids in atherosclerotic cardiovascular disease is punctuated by a lack of understanding regarding the metabolic fate and potential physiological effects of dietary peroxidized lipids and their decomposition products. This study sought to determine the metabolic fate and physiological ramifications of 13-hydroperoxyoctadecadienoic acid (13-HPODE) and 13-HODE (13-hydroxyoctadecadienoic acid) supplementation in intestinal and hepatic cell lines, as well as any effects resulting from 13-HPODE or 13-HODE degradation products. In the presence of Caco-2 cells, 13-HPODE was rapidly reduced to 13-HODE. Upon entering the cell, 13-HODE appears to undergo decomposition, followed by esterification. Moreover, 13-HPODE undergoes autodecomposition to produce aldehydes such as 9-oxononanoic acid (9-ONA). Results indicate that 9-ONA was oxidized to azelaic acid (AzA) rapidly in cell culture media, but AzA was poorly absorbed by intestinal cells and remained detectable in cell culture media for up to 18 h. An increased apolipoprotein A1 (ApoA1) secretion was observed in Caco-2 cells in the presence of 13-HPODE, 9-ONA, and AzA, whereas such induction was not observed in HepG2 cells. However, 13-HPODE treatments suppressed paraoxonase 1 (PON1) activity, suggesting the induction of ApoA1 secretion by 13-HPODE may not represent functional high-density lipoprotein (HDL) capable of reducing oxidative stress. Alternatively, AzA induced both ApoA1 secretion and PON1 activity while suppressing ApoB secretion in differentiated Caco-2 cells but not in HepG2. These results suggest oxidation of 9-ONA to AzA might be an important phenomenon, resulting in the accumulation of potentially beneficial dietary peroxidized lipid-derived aldehydes.

Synthesis and action of nitric oxide in rat glomerular mesangial cells

1991 ◽  
Vol 261 (4) ◽  
pp. F600-F606 ◽  
Author(s):  
P. J. Shultz ◽  
M. A. Tayeh ◽  
M. A. Marletta ◽  
L. Raij
Keyword(s):  
Nitric Oxide ◽  
Culture Medium ◽  
Mesangial Cells ◽  
Culture Media ◽  
Degradation Products ◽  
Specific Inhibitor ◽  
Cgmp Level ◽  
Glomerular Mesangial Cells ◽  
Intracellular Cgmp ◽  
Stimulation Of

Macrophages and certain tumor cell lines can be induced to synthesize nitric oxide (NO) from L-arginine after stimulation with lipopolysaccharide (LPS) or cytokines. In the present study, we have found that culture medium collected after 24 h from unstimulated rat mesangial cells (MC) contains 6.3 +/- 1.2 microM of NO3-/NO2- (the degradation products of NO). These levels were significantly increased when MC were incubated with LPS (10 micrograms/ml) for 24 h (23.9 +/- 4.1, P less than 0.05). The specific inhibitor of NO synthesis, NG-monomethyl-L-arginine (L-NMMA) completely inhibited LPS-stimulated production of NO3-/NO2-, confirming that the NO3-/NO2- was derived from NO within the MC. Recent studies suggest that endothelium-derived relaxing factor (EDRF) produced by vascular endothelium is also NO, and we have previously shown that both EDRF and NO stimulate increases in MC guanosine 3',5'-cyclic monophosphate (cGMP). Thus we sought to determine whether NO synthesized by the MC could affect cGMP levels within the same cells. After 24-h incubation with LPS (10 micrograms/ml), intracellular cGMP level within the MC was 706.3 +/- 197 (SE) compared with 40.5 +/- 7 fmol/micrograms protein in control MC incubated in media alone (P less than 0.01). The changes in cGMP in response to LPS were inhibited by greater than 90% by L-NMMA. Similar to LPS, incubation of MC with the cytokine gamma-interferon also increased NO3-/NO2- in the culture media and increased cGMP levels within MC. The induction of NO synthesis within MC and the concomitant stimulation of MC cGMP may be important in the modulation of the effects of endotoxemia, as well as inflammation, within the glomerulus.

scholarly journals Biosynthesis and distribution of leucocyte elastase inhibitor. Production of recombinant inhibitor.

1996 ◽  
Vol 43 (3) ◽  
pp. 497-501
Author(s):  
A Kasza ◽  
R Korpula-Mastalerz ◽  
S Rose-John ◽  
A Dubin
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Degradation Products ◽  
Interleukin 1 ◽  
Soluble Form ◽  
Kinetic Properties ◽  
Elastase Inhibitor ◽  
Elastin Degradation ◽  
Leucocyte Elastase ◽  
Marrow Cells

The horse leucocyte elastase inhibitor (HLEI), present in neutrophils, monocytes and bone marrow cells, is apparently a cytoplasmic protein which is not released from cells even in response to stimulation with lipopolysaccharide, phorbol ester, tumour necrosis factor alpha, interleukin-1 or elastin degradation products. Although no expression of the inhibitor was detected in neutrophils, both monocytes and bone marrow cells were efficient in its synthesis. Using a new expression vector pREST5d, recombinant inhibitor was produced in a large quantity in a soluble form, with a yield of 88 mg per 10 litres of E. coli culture. A two-step purification procedure, consisting of ion-exchange chromatography and gel filtration, yielded 36 mg of the recombinant inhibitor of a purity higher than 95%, as judged by SDS/PAGE. The recombinant protein had physicochemical and kinetic properties indistinguishable from those of the natural one, including irreversible elastase inhibition with an association rate constant kass > 10(7) M-1s-1. Both proteins were eliminated from rat circulation at the same ratio, and within the first 20 min 70% of the protein was removed. Such a short half-life in the circulation suggests that local delivery of HLEI directly to lungs in the form of aerosol could be a more efficient therapeutic approach than its intravenous injection.

scholarly journals The stereochemical course of reactions catalysed by the cellobiohydrolases produced by Talaromyces emersonii

1992 ◽  
Vol 283 (1) ◽  
pp. 31-34 ◽  
Author(s):  
M M Brooks ◽  
M G Tuohy ◽  
A V Savage ◽  
M Claeyssens ◽  
M P Coughlan
Keyword(s):  
Degradation Product ◽  
Proteolytic Degradation ◽  
Substrate Specificities ◽  
Talaromyces Emersonii ◽  
Anomeric Configuration ◽  
Culture Filtrates ◽  
Apparent Homogeneity ◽  
Displacement Reactions ◽  
Hydrolysis Of

Three forms of exocellobiohydrolase (EC 3.2.1.91), CBH IA, CBH IB and CBH II, were isolated to apparent homogeneity from culture filtrates of the aerobic fungus Talaromyces emersonii. CBH IA and CBH II appear to be native forms of these enzymes, while CBH IB may represent a proteolytic degradation product of the CBH IA enzyme. The hydrolysis of beta-cellobiosyl fluoride by each form was monitored by 1H-n.m.r. spectroscopy. The reactions catalysed by CBH IA and CBH IB proceed with retention of the anomeric configuration, whereas that catalysed by CBH II is one of inversion. Thus one may deduce that CBH IA (or CBH IB) and CBH II operate double and single displacement reactions respectively during catalysis of substrate. On the basis of these findings and the observed substrate specificities of the various forms, one may conclude that CBH IA (and CBH IB) is a family C enzyme, while CBH II belongs to family B [Henrissat, Claeyssens, Tomme, Lemesle & Mornon (1989) Gene 81, 83-95].

scholarly journals Identification of Immune Related LRR-Containing Genes in Maize (Zea maysL.) by Genome-Wide Sequence Analysis

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Wei Song ◽  
Baoqiang Wang ◽  
Xinghua Li ◽  
Jianfen Wei ◽  
Ling Chen ◽  
...  
Keyword(s):  
Zea Mays ◽  
Plant Disease Resistance ◽  
Interleukin 1 ◽  
Protein Structures ◽  
Gene Clusters ◽  
Amino Acid Sequences ◽  
Leucine Rich Repeat ◽  
Immune Receptors ◽  
Genome Wide ◽  
First Time

A large number of immune receptors consist of nucleotide binding site-leucine rich repeat (NBS-LRR) proteins and leucine rich repeat-receptor-like kinases (LRR-RLK) that play a crucial role in plant disease resistance. Although many NBS-LRR genes have been previously identified inZea mays, there are no reports on identifying NBS-LRR genes encoded in the N-terminal Toll/interleukin-1 receptor (TIR) motif and identifying genome-wide LRR-RLK genes. In the present study, 151 NBS-LRR genes and 226 LRR-RLK genes were identified after performing bioinformatics analysis of the entire maize genome. Of these identified genes, 64 NBS-LRR genes and four TIR-NBS-LRR genes were identified for the first time. The NBS-LRR genes are unevenly distributed on each chromosome with gene clusters located at the distal end of each chromosome, while LRR-RLK genes have a random chromosomal distribution with more paired genes. Additionally, six LRR-RLK/RLPs including FLS2, PSY1R, PSKR1, BIR1, SERK3, and Cf5 were characterized inZea maysfor the first time. Their predicted amino acid sequences have similar protein structures with their respective homologues in other plants, indicating that these maize LRR-RLK/RLPs have the same functions as their homologues act as immune receptors. The identified gene sequences would assist in the study of their functions in maize.

IMPURITY PROFILING IMPURITY PROFILING OF THIAMINE HYDROCHLORIDE INJECTION BY RP-HPLC AND CHARACTERIZATION OF DEGRADATION PRODUCT BY LC-MS/MS/QTOF

2020 ◽  
pp. 151-161
Author(s):  
SRINIVASU KONDRA ◽  
BAPUJI A. T. ◽  
D. GOWRI SANKAR ◽  
POTTURI MURALI KRISHNAM RAJU
Keyword(s):  
Degradation Product ◽  
Degradation Products ◽  
Hplc Method ◽  
Thiamine Hydrochloride ◽  
Injectable Formulation ◽  
Hyphenated Technique ◽  
Isolation And Characterization ◽  
Impurity Profiling ◽  
Rp Hplc

Objective: To propose a comprehensive, simple, and affordable RP-HPLC method for impurity profiling and characterization of unknown degradation products of thiamine hydrochloride injectable formulation. Methods: The chromatographic separation employs gradient mode using the octadecyl silane column using a mobile phase consisting of phosphate buffer with ion pair reagent, acetonitrile, and methanol delivered flow rate at 1.2 ml/min. The detection was carried out at 248 nm using empower software. LC-MS/MS/QTOF hyphenated technique was used for isolation and characterization of unknown degradation impurity. The performance of the method was systematically validated as per ICH Q2 (R1) guidelines. Results: Degradation product observed in accelerated stability was characterized by LC-MS/MS/QTOF hyphenated technique and found m/z value 351.1604 and postulated as an oxidative degradation product of thiamine due to excipient interaction. The validated method was sensitive, selective, and specific data proves the method is precise and accurate from LOQ to 150% level and results are within 95-108% and less than 4.5% RSD. The developed method is linear from 0.03-58.83 µg/ml with a correlation coefficient of more than 0.990 and LOD and LOQ value ranged from 0.03 to1.51 μg/ml. Conclusion: An efficient RP-HPLC method for impurity profiling of thiamine injectable formulation was successfully developed and unknown degradation product observed instability condition samples characterized by LC-MS/MS/QTOF technique. The validated method can be successfully employed for the impurity profiling of thiamine injectable in the quality control department.

Sign in / Sign up

Export Citation Format

Share Document