Hydrophobic interaction chromatography of fibroblast proteoglycans

AbstractWe 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 (GdnHCI). 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 HCI. 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.