A parallel between two techniques of extraction of connective tissue macromolecules.
A method of extraction of the collagen and noncollagen proteins from deep dermis of young adult rabbits using a 0.1 M tartaric acid solution was set up. The tartaric acid extraction, together with the preliminary neutral salt extraction, solubilized 95% of the total collagen and 98% of the noncollagen proteins, far more than the 6 M guanidinium Cl solution used for comparison. Elastin was not extracted. Studies on the fibrillation of the extracted collagen in neutral solution at 25 degrees C or on the results of pepsin digestion in acidic solution at +4 degrees C showed that the tartaric acid-extracted collagen was in a nondenatured form, whereas that extracted by guanidinium Cl was largely denatured. Polyacrylamide gel electrophoresis (PAGE) indicated that most of the collagen was of type I and that many noncollagen proteins were present, mostly in the molecular weight range of 40 kDa. Bidimensional PAGE gave a reproducible pattern of these noncollagen proteins, showing that several additional proteins were present in tartaric acid extracts and not in guanidinium chloride extracts.
Bellon
G
G
Laboratory of Biochemistry, University of Reims, France.
Wegrowski
J
J
Perreau
C
C
Randoux
A
A
Borel
J P
JP
Malgras
A
A
Chastang
F
F
D000818:Animals; D003094:Collagen / Q000302:isolation & purification*; D003238:Connective Tissue / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D019791:Guanidine; D006146:Guanidines; D011489:Protein Denaturation; D011506:Proteins / Q000302:isolation & purification*; D011817:Rabbits; D013644:Tartrates
D006146:Guanidines; D011506:Proteins; D013644:Tartrates; D003094:Collagen; D019791:Guanidine; C029768:tartaric acid