Summary
A sodium dodecylsulfate-polyacrylamide electrophoretic method, which in contrast to other biochemical procedures, e. g. differential salt precipitation or ion exchange chromatography and molecular sieve chromatography, is applicable to smallest amounts of protein, is shown to be suitable for the determination of the collagen types from small skin samples, such as routine skin biopsies.
After urea extraction, the tissue samples are cleaved with cyanogen bromide. The resulting CNBr peptides derived from the different α-chains are resolved in 12% SDS-polyacrylamide gels. Densitometric profiles of the gel electrophoretic patterns correspond to the collagen type content of the tissue specimens.
Comparing fetal to adult skin, the higher content of type III collagen in the case of fetal skin can be demonstrated.
Zusammenfassung
Es wird eine Dodecylsulfat-Polyacrylamidgel-elektrophoretische Methode beschrieben, mit welcher der Gehalt an Typ I- und Typ III-Kollagen in kleinen Hautproben, z. B. Probebiopsien, bestimmt werden kann. Im Gegensatz zu anderen biochemischen Methoden, z. B. der Salzpräzipitation oder der Ionenaustausch- und Molekularsiebchromatographie, ist diese Methode zur Darstellung kleinster Peptidmengen geeignet.
Harnstoffextrahierte Gewebeproben werden mit Bromcyan abgebaut. Die hierbei freigesetzten, den einzelnen α-Ketten zugeordneten Bromcyan-Peptide werden in 12%igen Gelen aufgetrennt. Aus den densitometrischen Kurvenverläufen der gelelektrophoretischen Peptidverteilungsmuster wird auf den Kollagentypgehalt der untersuchten Gewebeproben geschlossen.
Beim Vergleich von Fötalhaut mit Erwachsenenhaut kann der höhere Typ III-Gehalt der Fötalhaut dargestellt werden.
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Weber, L., Meigel, W.N. & Rauterberg, J. SDS-polyacrylamide gel electrophoretic. Arch. Derm. Res. 258, 251–257 (1977). https://doi.org/10.1007/BF00561127
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DOI: https://doi.org/10.1007/BF00561127