Hydrophobic interaction chromatography of fibroblast proteoglycans.
We have investigated the hydrophobic properties of human skin fibroblast proteoglycans and related material by affinity chromatography on Octyl-Sepharose CL-4B in 4 M guanidinium hydrochloride (GdnHCl). Proteoglycans and related material could be separated into non-, medium and highly hydrophobic forms by elution with gradients of Triton X-100 in 4 M Gdn HCl. The non-hydrophobic material included endogenously produced glycosaminoglycan chains and oligosaccharides as well as an HS-proteoglycan with a 35 kDa core. The 65-70 kDa core (glypican-related) proteoglycans appeared among the highly hydrophobic ones, but variable proportions were seen both in the medium and the non-hydrophobic material. Other membrane-bound proteoglycans, like fibroglycan (45 kDa core) and the HS-proteoglycans with 90 and 130 kDa cores, as well as the CS/DS-proteoglycan with a 90 kDa core, were all of high hydrophobicity. There were also indications of a highly hydrophobic CS/DS-proteoglycan with a 45 kDa core. The extracellular proteoglycans, PG-L, PG-S1 and PG-S2, and the HS-proteoglycans with 350 and 250 kDa cores were all of medium hydrophobicity. These proteoglycans emerged in distinct positions when the column was eluted with a gradient of 3-[(3-cholamidopropyl)dimethylammonio]propanesulphonate.
Schmidtchen
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A
Department of Physiological Chemistry, University of Lund, Sweden.
Fransson
L A
LA
D002478:Cells, Cultured; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002793:Cholic Acids; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000737:chemistry*; D006801:Humans; D017830:Octoxynol; D011092:Polyethylene Glycols; D011509:Proteoglycans / Q000737:chemistry* / Q000302:isolation & purification; D012685:Sepharose / Q000031:analogs & derivatives
D002793:Cholic Acids; D011509:Proteoglycans; D011092:Polyethylene Glycols; C027176:octyl-sepharose CL-4B; D017830:Octoxynol; D012685:Sepharose; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate