Publications [#113119] of Harold P. Erickson

Papers Published

1. HC Taylor, VA Lightner, WF Beyer, D McCaslin, G Briscoe, HP Erickson, Biochemical and structural studies of tenascin/hexabrachion proteins., Journal of cellular biochemistry, UNITED STATES, vol. 41 no. 2 (October, 1989), pp. 71-90, ISSN 0730-2312
   (last updated on 2009/02/12)

   Abstract:
   Tenascin is a large, disulfide-bonded glycoprotein of the extracellular matrix. The predominant form of tenascin observed by electron microscopy is a six-armed oligomer, termed a hexabrachion. We have determined the molecular mass of the native human hexabrachion to be 1.9 x 10(6) Da by sedimentation equilibrium analysis and by electrophoresis on non-reducing agarose gels. On reducing polyacrylamide gel electrophoresis (SDS-PAGE), human tenascin showed a single prominent band at 320 kDa and minor bands of 220 and 230 kDa. The molecular weight of the native human hexabrachion is thus consistent with a disulfide-bonded hexamer of the 320 kDa subunits. Upon treatment with neuraminidase, the apparent molecular weights of all human and chicken tenascin subunits on reducing SDS-PAGE were decreased by about 10 kDa. Prolonged incubation with alpha-mannosidase, however, caused no apparent change in the apparent molecular weight of tenascin subunits. Sedimentation in a cesium chloride gradient gave a higher buoyant density for human tenascin than for fibronectin, suggesting that it has a higher degree of glycosylation. The far-UV circular dichroism spectrum indicates a predominance of beta-structure and a lack of collagen-like or alpha-helical structure. When human hexabrachions were reduced and acetylated, the resulting fragments were single arms which sedimented at 6 S in glycerol gradients and migrated at 320 kDa on non-reducing gels. Treatment of tenascin with trypsin and alpha-chymotrypsin also produced large fragments which were fractionated by gradient sedimentation and analyzed by non-reducing SDS-PAGE and electron microscopy. We present a structural model for the assembly of the observed fragments into the elaborate native hexabrachion.

   Keywords:
   Animals • Cell Adhesion Molecules, Neuronal • Cells, Cultured • Chick Embryo • Circular Dichroism • Extracellular Matrix* • Fibroblasts • Glioma • Glycoside Hydrolases • Humans • Microscopy, Electron • Molecular Weight • Oxidation-Reduction • Protein Conformation • Tenascin • Tumor Cells, Cultured • analysis • isolation & purification • metabolism • ultrastructure*