The Isolation and Primary Structure of a 22-kDa Extracellular Matrix Protein from Bovine Skin

Abstract

The primary structure of a 22-kDa protein which was isolated during the purification of bovine skin dermatan sulfate proteoglycan is described. The uronate-rich fraction from DEAE-Sepharose chromatography of a 7.8 M urea extract of bovine fetal skin was subjected to gel filtration on Sepharose CL-6B in 4 M guanidine HCl. A prominent component of mass 22 kDa was separated from the proteoglycan and further purified on octyl-Sepharose. The primary structure of this component was determined and found to contain three repeat regions. Each of the three sections contains a similar pattern of looped disulfide bonds. A six-amino acid consensus sequence, Asp-Arg-Glx-Trp-Asn/Gln/Lys-Phe/Tyr, is found in each loop. This domain may be involved in associations of the molecule with other extracellular matrix components.