Premium
Affinity Purification of Proteoglycans that Bind to the Amyloid Protein Precursor of Alzheimer's Disease
Author(s) -
Williamson Timothy G.,
Nurcombe Victor,
Beyreuther Konrad,
Masters Colin L.,
Small David H.
Publication year - 1995
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1995.65052201.x
Subject(s) - proteoglycan , chondroitin sulfate proteoglycan , affinity chromatography , heparan sulfate , chemistry , neurite , biochemistry , perlecan , chondroitin sulfate , amyloid precursor protein , p3 peptide , glycosaminoglycan , extracellular matrix , alzheimer's disease , in vitro , enzyme , medicine , disease , pathology
The binding of the amyloid protein precursor (APP) to heparan sulfate proteoglycans has been shown to stimulate the neurite‐promoting activity of APP. In this study, proteoglycans that bind with high affinity to APP were characterized. Conditioned medium from cultures of postnatal day 3 mouse brain cells was applied to an affinity column containing a peptide homologous to a heparin‐binding domain of APP. A fraction 17‐fold enriched in proteoglycans was recovered by elution with a salt gradient. APP bound saturably and with high affinity to the affinity‐purified proteoglycan fraction. Scatchard analysis of the binding showed that APP bound to high‐ and low‐affinity sites with equilibrium dissociation constants of 1.4 × 10 −11 and 6.5 × 10 −10 M , respectively. APP, in conjunction with the affinity‐purified proteoglycan fraction, promoted neurite outgrowth. The affinity‐purified proteoglycan fraction contained a heparan sulfate proteoglycan and a chondroitin sulfate proteoglycan. Digestion of the affinity‐purified fraction with heparitinase I revealed a core protein of 63–69‐kDa molecular mass, whereas digestion with chondroitinase ABC revealed a core protein of 100–110 kDa. The results suggest that expression of specific APP‐binding proteoglycans may be an important step in the regulation of the neurite outgrowth‐promoting activity of APP.