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J F Woessner

Publications and source records attributed to J F Woessner.

At least 37 records · Page 2Linked to original sources
PubMed1995

Characterization of rat uterine matrilysin and its cDNA. Relationship to human pump-1 and activation of procollagenases.

A small uterine metalloproteinase of the rat has been shown by amino acid and cDNA sequencing to be orthologous to human pump-1. Both proteinases are now designated as matrilysin or matrix metalloproteinase 7. The properties of purified uterine metalloproteinase and recombinant pump-1 were compared. Their specificities on substrates (gelatins, fibronectin, transferrin, elastin, Azocoll, and (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3,[2, 4-dinitrophenyl]-L-2, 3-diaminopropionyl)-Ala-Arg-NH2) are similar and distinct from those of the stromelysins and gelatinases. The two matrilysins have similar sensitivity to hydroxamate and pseudopeptide inhibitors. Rat matrilysin selectively cleaves the alpha 2(I) chain of rat gelatin, producing major cuts at Gly713-decreases-Ile714, Gly775-decreases-Leu776, and Gly809-decreases-Ile810. Rat matrilysin produces maximum activation of latent human interstitial collagenase 1 (pro-matrix metalloproteinase 1) when added in the presence of 4-aminophenylmercuric acetate (APMA) by cleaving the Gln80-decreases-Phe81 bond. Rat and human matrilysin do not directly activate latent rat collagenase 3 (matrix metalloproteinase 13) and do not enhance its activation when added together with APMA. Autoactivation of collagenase 3 in the presence of APMA results in cleavage at Val81-decreases-Tyr82 corresponding to the Gln80-decreases-Phe81 cleavage in collagenase 1. Thus collagenase 3 is capable of maximal autoactivation, whereas collagenase 1 is dependent upon another matrix metalloproteinase in order to be activated to its full potential.

Amino Acid Sequence↗
PubMed1995

Matrilysin.

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Amino Acid Sequence↗
PubMed1994

Intraarticular injections with methylprednisolone acetate reduce osteoarthritic lesions in parallel with chondrocyte stromelysin synthesis in experimental osteoarthritis.

OBJECTIVE: To examine the effect of intraarticular injections of methylprednisolone acetate (MA) on osteoarthritic lesions and chondrocyte stromelysin synthesis in experimental osteoarthritis (OA). METHODS: In 15 mongrel dogs, the anterior cruciate ligament of the right knee was sectioned by a stab wound. Eight dogs received intraarticular injections of MA (20 mg) at the time of surgery and 4 weeks later; 7 had no treatment. The dogs were killed 8 weeks after surgery. Five normal dogs were used as controls. Macroscopic evaluation of the lesions, including measurements of osteophytes and areas of surface lesions on the condyles and plateaus, was conducted, along with histologic evaluation of the severity of lesions. Immunohistochemical analysis was carried out using a rabbit polyclonal antibody against stromelysin, followed by evaluation of matrix and chondrocyte staining using morphometric analysis. RESULTS: Treatment with MA significantly reduced the incidence (P < 0.0004) and size (P < 0.0001) of osteophytes. The histologic grading of cartilage lesions was also significantly reduced both on condyles (P < 0.01) and on plateaus (P < 0.002). Immunohistochemical studies revealed, for OA cartilage, a marked increase (P < 0.002) in the percentage of chondrocytes positive for stromelysin and in the intensity of staining throughout all the layers of the cartilage, as well as specific matrix staining (P < 0.005). Treatment with MA reduced staining at both the chondrocyte (P < 0.002) and the matrix (P < 0.01) levels toward normal. CONCLUSION: These findings provide additional evidence for the protective effect of corticosteroid injections on OA lesions, and indicate that the effect of this drug may be mediated through the suppression of stromelysin synthesis.

Animals↗
PubMed1993

Matrix metalloproteinase-3 (stromelysin-1). Identification as the cartilage acid metalloprotease and effect of pH on catalytic properties and calcium affinity.

Human pro-MMP-3 (pro-matrix metalloproteinase-3) was purified from three sources: articular cartilage and conditioned media from synovial fibroblasts and Chinese hamster ovary cells expressing recombinant pro-MMP-3. All three preparations reacted with two monoclonal antibodies specific for human fibroblast pro-MMP-3. Each preparation of active MMP-3 possessed properties identical to those previously reported for the cartilage acid metalloproteinase (MMP-6; Azzo and Woessner, J. F., Jr. (1986) J. Biol. Chem. 261, 5434-5441): an acid pH optimum of 5.3-5.5 for digestion of cartilage aggrecan; digestion of oxidized insulin B-chain at Ala14-Leu15 and Tyr16-Leu17 in a ratio of 3:1; and heat stability at neutral pH. Further characterization of MMP-3 establishes that the acid pH optimum for cartilage aggrecan is not due to substrate denaturation since the same optimum is found by viscosity assay, by SDS-polyacrylamide gel electrophoresis assay of G1 domain, and by digestion of aggrecan in fresh cartilage fragments in vitro. Fibronectin was also digested optimally at pH 5.5 and NH2-terminal sequence analysis revealed no pH change in a major proteolytic site of cleavage at the Pro689-Leu690 bond. The specificity constant kcat/Km is maximal at pH 5.5 as determined in a quenched fluorescence peptide assay. This is due to an increase in kcat at pH 5.5 without any substantial effect on Km. The affinity of MMP-3 for calcium is decreased about 10-fold at pH 5.3 compared to neutral pH. Finally, the neutral cartilage metalloproteinase is identified as 72-kDa pro-MMP-2 based on M(r), specificity of insulin B-chain cleavage, and reactivity with a specific polyclonal antibody to human MMP-2.

Amino Acid Sequence↗
PubMed1993

Activation of cartilage stromelysin-1 at acid pH and its relation to enzyme pH optimum and osteoarthritis.

Stromelysin-1 was purified from human articular cartilage and compared to synovial fibroblast enzyme and to recombinant enzyme. If the latent enzyme was incubated at pH 5.5 with substrates such as aggrecan, it spontaneously became active. Incubation of latent zymogen alone at pH 5.5 gave increasing activation over a period of at least 5 hours. However, this activation process was not accompanied by any shift in molecular weight even after continued incubation for 18 hours. Maximum activity observed was 45-60% of that seen with APMA activation. Stromelysin-1 has a pH optimum of 5.5-6.5 on various macromolecular and peptide substrates. Interaction with TIMP is reduced at pH 5.5 relative to that at 7.5. The hypothesis is presented that osteoarthritis may be initiated by acid activation of stromelysin-1.

Cartilage, Articular↗
PubMed1993

Collagenase, its inhibitors, and decorin in the lower uterine segment in pregnant women.

OBJECTIVE: It is postulated that collagenase activity in the cervix and lower uterine segment is important for dilatation at the time of labor. If so, the enzyme must partially escape from control by inhibitors. A second hypothesis is that an elevated ratio of decorin (dermatan sulfate proteoglycan) to collagen may also contribute to the dilatation process. STUDY DESIGN: The activity of collagenase, its natural inhibitors (tissue inhibitor of metalloproteinase-1 and alpha 2-macroglobulin), elastase, hydroxyproline (collagen), and decorin were quantified in biopsies of the lower uterine segment at term (not dilated) and during active labor. RESULTS: Collagenase concentration is 23 times higher in the cervix of patients in labor than at term. The sum of alpha 2-macroglobulin plus tissue inhibitor of metalloproteinase-1 increased only twofold in labor, as did elastase. The ratio of decorin to collagen doubled. CONCLUSION: The imbalance caused by collagenase increasing much more than its inhibitors may contribute to collagen breakdown and dilatation. Neutrophils may be responsible for much of this increase of collagenase. The increased ratio of decorin to collagen supports the hypothesis that the interaction of these two components is important in dilatation.

Adult↗
PubMed1993

The angiotensin II antagonist saralasin inhibits ovulation in the perfused rat ovary.

OBJECTIVE: Our null hypothesis was that the angiotensin II antagonist saralasin does not reduce the number of ovulations in the rat ovarian perfusion model. STUDY DESIGN: Ovaries from pregnant mare's serum gonadotropin-stimulated immature rats were perfused with nutrient media to which luteinizing hormone and 3-isobutyl-1-methylxanthine had been added to induce ovulation. Test perfusions were treated with saralasin 1 mumol/L (n = 0.5) and compared with controls (n = 5) with the Student t test. Perfusions with both saralasin and angiotensin II and dose-response evaluations were performed. RESULTS: Saralasin-treated ovulations were 6.6 +/- 1.3 (mean + SEM) compared with 18.6 +/- 3.9, p < 0.02. The effects of saralasin could be reversed with the addition of an equimolar amount of angiotensin II. Dose-response evaluations showed a progressive inhibition of ovulation at 10(-8) to 10(-6) mol/L. CONCLUSION: The angiotensin II antagonist saralasin inhibits ovulation in a dose-dependent fashion; this effect is canceled by the addition of equimolar concentrations of angiotensin II.

Angiotensin II↗
PubMed1992

cDNA sequence for rat dermatan sulfate proteoglycan-II (decorin).

A cDNA clone for dermatan sulfate proteoglycan-II, or decorin, has been isolated from a rat uterus library and sequenced. The cDNA and deduced amino acid sequences are 79 and 77% identical to the previously reported human and bovine sequences, respectively. The rat protein contains potential attachment sites for two glycosaminoglycan chains and four N-linked oligosaccharides, six conserved cysteine residues and multiple repeats of a leucine-rich sequence, LXXLXLXXNXL/I. Overlapping the C-end of one of these repeats is an NKISK sequence, which has been implicated in binding to fibronectin.

Amino Acid Sequence↗
PubMed1992

Serum collagenase levels in relation to the state of the human cervix during pregnancy and labor.

OBJECTIVE: The purpose of our study was to investigate the role of collagenase in the cervical ripening and dilatation process in term pregnancy. STUDY DESIGN: Serum samples were obtained from nonpregnant women (n = 5) and term-pregnant women. The term-pregnant women were either admitted for elective cesarean section or labor induction (unfavorable cervix, n = 19; favorable cervix, n = 12) or in spontaneous, active labor (stiff and inelastic cervix, n = 7; soft compliant cervix, n = 8). Statistical analysis was performed with the Student t test. RESULTS: The nonpregnant women had low serum collagenase levels (5.2 +/- 0.7 micrograms collagen digested per minute per 100 ml serum, mean +/- SEM). At term but before labor, women with unripe cervices had higher collagenase levels (10.3 +/- 0.9). The women with ripe cervices had even higher serum collagenase levels (22.9 +/- 4.2; p < 0.001). During labor, women with stiff and inelastic cervices had lower serum collagenase levels compared with women with soft and compliant cervices (12.9 +/- 1.7 vs 28.0 +/- 4.2; p < 0.01). CONCLUSION: Serum collagenase during ripening at term and in active labor increases, supporting its active role in the ripening process.

Adult↗
PubMed1992

Collagenase in wound healing: effect of wound age and type.

Collagenase is believed to be important for cell migration and collagen remodeling during tissue repair and regeneration. We have investigated collagenase concentrations in different types of surgically inflicted wounds in pigs. Collagenase was extracted from tissue homogenates of wounds by heating to 60 degrees C for 6 min in 0.1 M CaCl2. The molecular weight of latent collagenase was about 52 kDa. Activated collagenase produced the characteristic 3/4 fragment of collagen. Collagenase was assayed by the use of radiolabeled telopeptide-free collagen. To detect maximal collagenase activity, extracts were reduced and alkylated to destroy inhibitors, then activated with aminophenylmercuric acetate. Sutured incisions showed peak collagenase content on postoperative day 1 and thereafter steadily declining concentrations. Granulation tissue from non-sutured large defect full-thickness wounds showed high collagenase content on postoperative day 5 and then a sharp decline to day 7 followed by a slowly declining curve to postoperative day 21. Partial-thickness wounds exhibited a different time course, with collagenase increasing to peak concentrations on postoperative days 3-5; however, a large proportion of the detected collagenase was due to the adherent scab. By day 7 collagenase concentrations approached the low concentrations of normal skin when epithelialization was complete and the scab rejected. In general, collagenase shows an early maximum and then declines with postoperative time, with the sharpest decline occurring when epithelialization is complete.

Aging↗