Anchorage of collagen-tailed acetylcholinesterase to the extracellular matrix is mediated by heparan sulfate proteoglycans.
Heparan sulfate and heparin, two sulfated glycosaminoglycans (GAGs), extracted collagen-tailed acetylcholinesterase (AChE) from the extracellular matrix (ECM) of the electric organ of Discopyge tschudii. The effect of heparan sulfate and heparin was abolished by protamine; other GAGs could not extract the esterase. The solubilization of the asymmetric AChE apparently occurs through the formation of a soluble AChE-GAG complex of 30S. Heparitinase treatment but not chondroitinase ABC treatment of the ECM released asymmetric AChE forms. This provides direct evidence for the vivo interaction between asymmetric AChE and heparan sulfate residues of the ECM. Biochemical analysis of the electric organ ECM showed that sulfated GAGs bound to proteoglycans account for 5% of the total basal lamina. Approximately 20% of the total GAGs were susceptible to heparitinase or nitrous acid oxidation which degrades specifically heparan sulfates, and approximately 80% were susceptible to digestion with chondroitinase ABC, which degrades chondroitin-4 and -6 sulfates and dermatan sulfate. Our experiments provide evidence that asymmetric AChE and carbohydrate components of proteoglycans are associated in the ECM; they also indicate that a heparan sulfate proteoglycan is involved in the anchorage of the collagen-tailed AChE to the synaptic basal lamina.
Brandan
E
E
Maldonado
M
M
Garrido
J
J
Inestrosa
N C
NC
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism; D004557:Electric Organ / Q000201:enzymology* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000201:enzymology*; D005399:Fishes; D006025:Glycosaminoglycans / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000201:enzymology* / Q000648:ultrastructure; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism*
D000110:Acetylcholinesterase; D011508:Chondroitin Sulfate Proteoglycans; D003094:Collagen; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006493:Heparin; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D011509:Proteoglycans; C021349:heparitinsulfate lyase