The chemistry of connective tissues. 3. Composition of the soluble proteins derived from elastin

Abstract

Vol. 6i PARTIAL HYDROLYSIS OF ELASTIN 21 p-proteins were separated by repeated precipitation of the coacervate at 600; osmotic-pressure determinations on the preparations gave a mean molecular weight of 67000 for the a-protein and 5500 for the p-protein.The a-protein had ash 0.03%. Analysis by combustion (Weiler and Strauss, Oxford) gave C, 52-9; H, 7-4; N, 16.5%. The ,-protein had ash 0-28; C, 52-4; H, 7-75; N, 16-6 % (analyses corrected for ash). METHODSAmino acid analy8i8 of ela8tin and the two derived protein8 The procedure adopted for the separation and estimation of the amino acids was that of . The two samples of elastin powder, and the standard preparations of a-and ,B-protein were hydrolysed with acid and chromatographed under identical conditions. For hydrolysis, the protein (500 mg.) was mixed with 6N-HC1 (50 ml.) and the mixture refluxed for 24 hr. Hydrochloric acid was then removed by repeated evaporation in vacuo, and the syrup made up to 50 ml. with distilled water. Portions were then taken for determination of N (Kjeldahl). Equal volumes of the hydrolysate and sodium citrate buffer (0.1 M, pH 3.42) were then mixed, and 1 ml. of the buffered solution was applied to the columns. The weight of protein equivalent to the amino acid mixture applied to the column was calculated from the nitrogen content of the hydrolysate. The management of the columns exactly followed the procedure given by .Arginine, lysine and histidine were estimated using the short (15 cm.) column of Moore & Stein. Hydroxyproline was estimated in separate experiments in which the 100 cm. column was first equilibrated with a sodium citrate buffer of slightly lower pH (pH 3.04). The column was then developed with a further quantity of pH 3-04

Interaction of l-lysine and soluble elastin with the semicarbazide-sensitive amine oxidase in the context of its vascular-adhesion and tissue maturation functions

“…Some small lysine-containing peptides have been reported to to inhibit SSAO [31,32]. Since the elastin is rich in lysine [19,33], and many have reported on the role of SSAO in the maturation of the ECM [14,34] we investigated the possible effects of soluble elastin on the activity of SSAO. The peptide was found to cause inhibition of the initial rates of oxidation of methylamine and benzylamine by the enzyme.…”

“…For binding assays a preparation of SSAO by the supplier BioVar Ltd, Yerevan, Armenia, was also used. L-lysine, poly-L-lysine hydrobromide (M r range =1000 to 5000) and soluble elastin, purified from bovine neck ligament by the method of Partridge et al [18,19], were purchased from Sigma-Aldrich. Amplex Red reagent was purchased from Invitrogen.…”

Processing and characterization of α-elastin electrospun membranes

“…The partial hydrolysis of elastin was based on the procedure described by Partridge et al 24 . Elastin was treated with 0.25 M oxalic acid (ratio of 1 g elastin: 7.5 ml acid) at 100 ºC for 1 hour with vigorous stirring.…”

The Use of Natural Polymers in Tissue Engineering: A Focus on Electrospun Extracellular Matrix Analogues

“…The incorporation of elastin as a biomaterial has been used in several forms, including insoluble elastin in autografts, allografts, xenografts, decellularized ECM, and in purified elastin forms where the insoluble elastin has been hydrolyzed to a soluble form. Repeated elastin-like sequences have been produced by synthetic or recombinant means [73], and even recombinant tropoelastin or tropoelastin fragments have been used as biomaterials [74,75]. The use of soluble elastin is advantageous over its insoluble form due to the straightforwardness in the handling and analysis of the material.…”