Detergent-solubilized proteoglycans in rat testicular Sertoli cells.
Rat Sertoli cells were cultured for 48 h in the presence of [35S]sulfate and extracted with 4 M guanidine chloride. In this extract, a Sepharose CL-2B Kav 0.10 proteoheparan appeared lipid associated, since after addition of detergent it emerged at Kav = 0.65 on Sepharose CL-2B. Treatment of cells with 0.2% Triton X-100 released 35S-labeled material which was purified by ion-exchange chromatography and hydrophobic interaction chromatography on octyl-Sepharose. Proteoglycan with affinity for octyl-Sepharose (Kav = 0.30 and 0.12 on Sepharose CL-4B and CL-6B, respectively) mostly carried heparan sulfate chains with Kav = 0.38 and minor proportion of heparan chains with Kav = 0.77 on Sepharose CL-6B. An association with lipids was confirmed by intercalation into liposomes of this proteoheparan which might be anchored in the plasma membrane, via an hydrophobic segment and/or covalently linked to an inositol-containing phospholipid. Non-hydrophobic material consisted of: (i) proteoheparan slightly smaller in size than lipophilic proteoheparan and possibly deriving from this one and (ii) two heparan sulfate glycosaminoglycan populations (Kav = 0.38 and 0.86 on Sepharose CL-6B) corresponding to single glycosaminoglycan chains and their degradation products.
Mounis
A
A
Laboratoire de Biochimie, URA CNRS 609, IBBA, Université de Caen Esplanade de la Paix, France.
Barbey
P
P
Langris
M
M
Bocquet
J
J
D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003902:Detergents; D008297:Male; D017830:Octoxynol; D011092:Polyethylene Glycols; D011509:Proteoglycans / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012708:Sertoli Cells / Q000737:chemistry*; D012995:Solubility; D013737:Testis / Q000737:chemistry*
D003902:Detergents; D011509:Proteoglycans; D011092:Polyethylene Glycols; D017830:Octoxynol