| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1986: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1985: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
We have isolated a glycopeptide named nephritogenoside, which was purified from the glomerular basement membrane of rats. Nephritogenoside contains a new type of carbohydrate-peptide linkage having the <alpha> -D-configuration. After the cleavage of the amido-linkage of nephritogenoside between carbohydrate and peptide by ion-exchange resin, 1N-hydrochloric acid eluate was chromatographed on a column of Sephadex G-100 to give a single peptide peak (Fr. 10). The amino acid sequence of Fr. 10 has been determined with an amino acid s quencer. Based on the results of the experiment in which nephritogenoside was treated with carboxypeptidase Y, we have established that the trisaccharide chain and the peptide portion bind directly and N- <alpha> -glycosidically with N-terminal aspartic acid, as represented by following structure, Glc <alpha> -(1->6)Glc <beta> -(1->6)Glc- <alpha> -Asn-Pro- Leu-Phe-Gly-Ile-Ala-Gly-Glu-Asp-Gly-Pro-Thr-Gly-Pro-Ser-Gly-Ile-Val-Gly-Gln.
O-( <alpha> -D-Glucopyranosyl)-(1->6)-O- <beta> -D-glucopyranosyl)- (1->6)-1-N-(Asp-Pro)-, -Asp-Pro-Leu-Phe-Gly)-, -Asp-Pro-Leu-Phe-Gly-Ile-Ala-Gly)- <alpha> -D-glucopyranosylamines have been prepared, as a model of a derivative of nephritogenoside, by condensation of the trisaccharide amine with the corresponding of the peptide derivatives in the presence of DEPC-TEA. Protective groups were removed by O-deacylation, and hydrogenolysis. The aspartyl derivative of the trisaccharide was also converted into the acyl azide which was condensed with bovine serum albumin to form a neoglycoprotein.
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