The chemistry of connective tissues. 2. Soluble proteins derived from partial hydrolysis of elastin

Partridge, S. M.; Davis, H. F.; Adair, G. S.

doi:10.1042/bj0610011

Cited by 466 publications

(79 citation statements)

References 17 publications

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“…TE concentration was determined by the method of ELISA as described before (12) using chicken anti-rat elastin antibody (1 :2000), and rat a-elastin (40 ng/ 100 pL/well) was purified from rat aorta by the method of Partridge et al (16) The fibroblastic characteristics of cells were examined to determine if the cells had any smooth muscle-like characteristics. Ultrastructurally, the cells have cytoplasmic lipid vacuoles, a small amount of microfilaments, and prominent cisternae of rough endoplasmic reticulum.…”

Section: Total Protein Synthesis Protein Synthesis Was Estimated Bymentioning

confidence: 99%

IGF-I Stimulates Tropoelastin Synthesis in Neonatal Rat Pulmonary Fibroblasts

Noguchi

Nelson

1991

Pediatr Res

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ABSTRACT. We have examined the effect of IGF-I on tropoelastin (TE) synthesis in cultured rat neonatal pulmonary fibroblasts, because this growth factor has been shown to stimulate TE synthesis in vascular smooth muscle cells. IGF-I stimulated TE and total protein synthesis in a dose-dependent manner even when cells were cultured in the medium supplemented with 0.5% FCS. The maximal stimulation was at IGF-I concentration 500 ng/mL and was an increase of 86 f 14 and 35 f 5% for TE and estimated total protein synthesis, respectively. There was a corresponding 95 f 20% increase in the TE mRNA/fi-actin mRNA ratio assessed by densitometry of the Northern blot analysis. At this low concentration of FCS, however, there was neither TE stimulation by dexamethasone alone nor in combination with IGF-I. We conclude that IGF-I stimulation of TE synthesis may occur in cells other than vascular smooth muscle cells and that there is no additive stimulation by glucocorticoids. (Pediatr Res 30: 248-251, 1991) Abbreviations RPF, rat pulmonary fibroblasts TE, tropoelastin DEX, dexamethasone An extracellular protein component, elastin, may play an important role in the formation of the terminal airsac/alveolus during perinatal lung morphogenesis in various animals and in humans (1-8).The pro-form protein, TE, is a soluble protein of approximately 72 000 to 75 000 molecular weight (9, lo), and the regulation of synthesis is poorly understood, partly because regulatory elements of the gene have not been fully defined (1 1). We have previously demonstrated that cultured RPF are elastogenic in early passaged cells and that TE expression is stimulated by DEX (12). To further understand the humoral factors that could stimulate TE synthesis in the developing lung, we have examined IGF-I in the present study. IGF-I has been shown to stimulate TE expression in vascular smooth muscle tissue and cells (13,14), but this effect on other cell types including RPF has not been reported. We found that IGF-I stimulates TE synthesis in the neonatal RPF in a dose-dependent manner. MATERIALS AND METHODSapolis, IN). The project was approved by the Animal Care Committee of St. Louis University. Postnatal d 1 was counted as the birth date, and RPF were isolated as described before (12). Selection of the ages and response to DEX stimulation was dependent on our previous finding that TE expression is maximum at 7 d (12). Pups from two or three litters were combined and were used for extraction of total RNA and isolation of pulmonary fibroblasts by the differential adhesion method (1 5). The cells were passaged once and maintained in RPMI 1640 supplemented with 10% FCS until just before confluency and then in RPMI 1640 with 0.5% FCS to minimize the IGF-I effects of the FCS (14). At this FCS concentration, the results were more consistent than with 0.1 % FCS. Twelve h after the medium switch, cells were treated with increasing concentrations of IGF-I for another 48 h, harvested by 0.25% trypsin and 0.0 1 % EDTA treatment, and processed for extraction of tot...

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Section: Total Protein Synthesis Protein Synthesis Was Estimated Bymentioning

confidence: 99%

IGF-I Stimulates Tropoelastin Synthesis in Neonatal Rat Pulmonary Fibroblasts

Noguchi

Nelson

1991

Pediatr Res

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Section: Hydrolyzed Elastin Materialsmentioning

confidence: 99%

“…A fragmented elastin preparation termed -elastin is obtained by hydrolysis with oxalic acid and is often used in in vitro studies of elastin (Partridge, Davis et al 1955). Hydrolysis can be performed with potassium hydroxide to yield κ-elastin or through mild digestion with proteinases (Partridge, Davis et al 1955;Jacob and Hornebeck 1985). Hydrolyzed preparations of elastin display various properties that are similar to the native protein including temperature-induced aggregation (coacervation) and regulation of SMC and fibroblast phenotype (De Vries, Zeegelaar et al 1995;Ito, Ishimaru et al 1998).…”

Section: Hydrolyzed Elastin Materialsmentioning

confidence: 99%

Elastin Based Constructs

Nivison‐Smith¹,

Weiss²

2011

Regenerative Medicine and Tissue Engineering - Cells and Biomaterials

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“…Its purity was checked by amino acid analysis. Soluble elastin peptides were obtained by partial hydrolysis of highly purified insoluble elastin with oxalic acid (11). A peptide with an average molecular mass of 25 kDa was isolated by column chromatography on Sephadex G75.…”

Section: Methodsmentioning

confidence: 99%

Elastin‐derived peptide induces monocyte chemotaxis by increasing intracellular cyclic GMP level and activating cyclic GMP dependent protein kinase

Uemura¹,

Okamoto²

1997

IUBMB Life

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SUMMARY: An elastin-derived peptide with an average molecular mass of 25 kDa was shown to induce monocyte chemotaxis at the optimal concentration of 10 -1 o,g/ml. Homologous deactivation test showed that monocytes exposed to the elastin-derived peptide at 10 "1 wg/ml lost their chemotactic responsiveness when reexposed to the same stimulus. In conjunction with chemotactic response to the elastin-derived peptide, intracellular guanosine 3', 5'-monophosphate (cGMP) levels were enhanced but intracellular adenosine 3', 5'-monophosphate (cAMP) levels were not. The monocyte migration induced by the elastinderived peptide was inhibited by cGMP dependent protein kinase (PKG) inhibitor, but not by cAMP dependent protein kinase inhibitor and protein kinase C inhibitor. These results suggest that the elastin-derived peptide induces monocyte chemotaxis by increasing the level of cGMP, followed by activating PKG.

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The chemistry of connective tissues. 2. Soluble proteins derived from partial hydrolysis of elastin

Cited by 466 publications

References 17 publications

IGF-I Stimulates Tropoelastin Synthesis in Neonatal Rat Pulmonary Fibroblasts

IGF-I Stimulates Tropoelastin Synthesis in Neonatal Rat Pulmonary Fibroblasts

Elastin Based Constructs

Elastin‐derived peptide induces monocyte chemotaxis by increasing intracellular cyclic GMP level and activating cyclic GMP dependent protein kinase

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