Two Improved and Simplified Methods for the Spectrophotometric Determination of Hydroxyproline.

Bergman, I.; Loxley, Roy

doi:10.1021/ac60205a053

Cited by 1,416 publications

(647 citation statements)

References 7 publications

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“…Hydroxyproline release was assayed (as a measure of collagen degradation) using a microtiter modification of the method as described previously (45,46).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of furin‐like enzymes blocks interleukin‐1α/oncostatin M–stimulated cartilage degradation

Milner¹,

Rowan²,

Sf³

et al. 2003

Arthritis & Rheumatism

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Objective. To investigate the role of furin-like enzymes in the proteolytic cascades leading to cartilage breakdown and to examine which collagenase(s) contribute to collagen degradation.Methods. Bovine nasal cartilage was stimulated to resorb with the addition of interleukin-1␣ (IL-1␣)/ oncostatin M (OSM) in the presence or absence of a furin inhibitor, Dec-RVKR-CH 2 Cl, or selective matrix metalloproteinase 1 (MMP-1) inhibitors. Collagen and proteoglycan levels were determined by assay of hydroxyproline and sulfated glycosaminoglycan, respectively. Collagenase and gelatinase activity were measured using 3 H-acetylated collagen and gelatin zymography, respectively.Results. The addition of Dec-RVKR-CH 2 Cl to stimulated cartilage reduced the release of collagen fragments and the levels of active collagenase and MMP-2, suggesting that furin-like enzymes are involved in the cascades leading to activation of procollagenases. At MMP inhibitor concentrations that selectively inhibit MMP-1, no inhibition of collagen release was observed, but increasing the concentration to the 50% inhibition concentration for MMP-13 resulted in a 50% blockage of collagen release. The addition of Dec-RVKR-CH 2 Cl to resorbing cartilage also partially blocked proteoglycan release, thus demonstrating a role for furin-activated enzymes in the pathways leading to proteoglycan degradation. Conclusion.Furin-like enzymes are involved in cascades leading to activation of procollagenases and degradation of collagen. MMP-13, which can be activated by furin-processed membrane-type 1 MMP-1, appears to be a major collagenase involved in collagen degradation induced by IL-1␣/OSM. Furin-like enzymes also appear to play a role in the pathways leading to proteoglycan degradation. These findings are of importance when considering proteinase inhibition as a target for therapeutic intervention in arthritic diseases.

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“…Hydroxyproline release was assayed (as a measure of collagen degradation) using a microtiter modification of the method as described previously (45,46).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of furin‐like enzymes blocks interleukin‐1α/oncostatin M–stimulated cartilage degradation

Milner¹,

Rowan²,

Sf³

et al. 2003

Arthritis & Rheumatism

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show abstract

“…Collagen measurements -The method of Bergman and Loxley (1963) was used to determine collagen concentration by the measurement of hydroxyproline content. Briefly, after fixation in neutral 10% formalin the snails were hydrolyzed for 9 to 18 h in hydrochloric acid at 110°C, neutralized in NaOH (10N) and HCl (6N).…”

Section: Methodsmentioning

confidence: 99%

Extra-cellular matrix changes in Schistosoma mansoni-infected Biomphalaria glabrata

Borges

Andrade

2003

Mem. Inst. Oswaldo Cruz

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“…Biochemical study -A sample of the liver was frozen at -70 o C for determination of collagen, measured as hydroxyproline by the spectrophotometric method B of Bergman and Loxley (1963). Hydroxyproline levels were corrected for intensity of infection with help of a simplified electronic spread sheet elaborated by Cheever (1986) and used in subsequent papers.…”

Section: Experimental Groupsmentioning

confidence: 99%

Malnutrition and hepatic fibrosis in murine schistosomiasis

Coutinho

2004

Mem. Inst. Oswaldo Cruz

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Two Improved and Simplified Methods for the Spectrophotometric Determination of Hydroxyproline.

Cited by 1,416 publications

References 7 publications

Inhibition of furin‐like enzymes blocks interleukin‐1α/oncostatin M–stimulated cartilage degradation

Inhibition of furin‐like enzymes blocks interleukin‐1α/oncostatin M–stimulated cartilage degradation

Extra-cellular matrix changes in Schistosoma mansoni-infected Biomphalaria glabrata

Malnutrition and hepatic fibrosis in murine schistosomiasis

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