1. Non-collagenous substances in newborn calf dermis were extracted with solutions of various concentrations of MgCl
2. The total protein and hydroxyproline contents in MgCl
2 extracts increased with increase in the concentration of MgCl
2 in the solutions. In particular, steep increases of their contents were observed at concen-trations of MgCl
2 from 0.5 to 1.0M. Total amounts of hydroxyproline in 1.0, 2.0, and 3.0M MgCl
2 extracts were equivalent to 40-50% of the hydroxyproline content in the whole connective tissue. Hexose and hexosamine contents of MgCl
2 extracts increased with increase of the MgCl
2 concentration. Hexuronic acid was hardly present in the residues after extractions with 0.5, 1.0, 2.0, and 3.0 M MgCl
2.
2. Plasma proteins, hyaluronic acid, and dermatan sulfate were extracted at low concentrations of MgCl
2. A
non-
collagenous protein and MgCl
2-soluble collagen were extracted with 1.0, 2.0, and 3.0M MgCl
2 solutions. The disperson of collagen fibrils was observed in the residue extracted with 1.0M MgCl
2 solution by electron micro-scopy; the fibril structure of collagen was disordered by extraction with 2.0 and 3.0M MgCl
2. These results suggest that the dispersion and disorder of collagen fibrils lead to the release of a
non-
collagenous protein. Furthermore, it is suggested that the removal of hyaluronic acid and dermatan sulfate was not very effective for the solubilization of a large amount of collagen, but was suitable as a pretreatment to the extraction of a
non-
collagenous protein accompanied by the solubilization of a large amount of collagen.
3. The
non-
collagenous protein was purified by DEAE-cellulose column chromatog-raphy. Polyacrylamide gel electrophoresis of this protein at pH 8.5 showed a single band moving to the cathode. The
non-
collagenous protein contained 3.7% hexose, 1.8% hexosamine, and no hexuronic acid. This protein is rich in glycine, glutamic acid, and alanine, and contains neither hydroxyproline nor hydroxylysine. Sedimen-tation analysis showed a single peak with 1.8S and the molecular weight was approx. 43, 000 as determined by SDS polyacrylamide gel electrophoresis.
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