弗氏小鼠白血病病毒糖蛋白71中各个糖基化位点的寡糖
Oligosaccharides at individual glycosylation sites in glycoprotein 71 of Friend murine leukemia virus.
作者信息
Geyer R, Dabrowski J, Dabrowski U, Linder D, Schlüter M, Schott H H, Stirm S
机构信息
Biochemisches Institut am Klinikum der Justus-Liebig-Universitat, Giessen, Federal Republic of Germany.
出版信息
Eur J Biochem. 1990 Jan 12;187(1):95-110. doi: 10.1111/j.1432-1033.1990.tb15281.x.
Glycoprotein 71 from Friend murine leukemia virus was digested with proteases and the glycopeptides obtained were isolated and assigned, by amino acid sequencing, to the eight N-glycosylated asparagines in the molecule; only Asn334 and Asn341 could not be separated. The oligosaccharides liberated from each glycopeptide by endo-beta-N-acetylglucosaminidase H, or by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F, were fractionated and subjected to structural analysis by one- and two-dimensional 1H NMR, as well as by methylation/gas-liquid-chromatography/mass-fragmentography. At each glycosylation site, the substituents were found to be heterogeneous including, at Asn334/341 and Asn410, substitution by different classes of N-glycans: oligomannosidic oligosaccharides, mainly Man alpha 1----6(Man alpha 1----3)Man alpha 1----6(Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4GlcNAc beta 1----, were detected at Asn168, Asn334/341 and Asn410. Hybrid species, partially sialylated, intersected and (proximally) funcosylated Man alpha 1----6(Man alpha 1----3)Man alpha 1----6 and Man alpha 1----3Man alpha 1----6 and Man alpha 1----3Man alpha 1----6(Gal beta 1----4GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4GlcNAc beta 1----, were found at Asn12, as previously published [Schlüter, M., Linder, D., Geyer, R., Hunsmann, H., Schneider, J. & Stirm, S. (1984) FEBS Lett. 169, 194-198] and at Asn334/341. N-Acetyllactosaminic glycans, mainly partially intersected and fucosylated NeuAc alpha 2----3 or Gal alpha 1----3Gal beta 1----4GlcNAc beta 1----2Man alpha 1----6(NeuAc alpha 2----6 or NeuAc alpha 2----3Gal-beta 1----4GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNac beta 1----4GlcNAc beta 1---- with some bifurcation at ----6Man alpha 1----6, were obtained from Asn266, Asn302, Asn334/341, Asn374 and Asn410. In addition, Thr268, Thr277, Thr279, Thr304/309, as well as Ser273 and Ser275, were found to be O-glycosidically substituted by Gal beta 1----3GalNAc alpha 1----, monosialylated or desialylated at position 3 of Gal or/and position 6 of GalNAc.
将来自弗氏鼠白血病病毒的糖蛋白71用蛋白酶消化，所得糖肽经分离后，通过氨基酸测序确定分子中8个N - 糖基化天冬酰胺的归属；只有Asn334和Asn341无法分离。用内切β - N - 乙酰葡糖胺酶H或肽 - N4 -（N - 乙酰 - β - 葡糖胺基）天冬酰胺酰胺酶F从每个糖肽中释放出的寡糖进行分级分离，并通过一维和二维1H NMR以及甲基化/气 - 相色谱/质谱 - 碎片谱进行结构分析。发现在每个糖基化位点，取代基都是异质的，在Asn334/341和Asn410处，被不同类型的N - 聚糖取代：寡甘露糖型寡糖，主要是Manα1→6(Manα1→3)Manα1→6(Manα1→3)Manβ1→4GlcNAcβ1→4GlcNAcβ1→，在Asn168、Asn334/341和Asn410处被检测到。部分唾液酸化、交叉连接且（近端）岩藻糖基化的杂合型Manα1→6(Manα1→3)Manα1→6和Manα1→3Manα1→6以及Manα1→3Manα1→6(Galβ1→4GlcNAcβ1→2Manα1→3)Manβ1→4GlcNAcβ1→4GlcNAcβ1→，在Asn12处被发现，如先前发表的[施吕特，M.，林德，D.，盖耶，R.，洪斯曼，H.，施奈德，J. & 施蒂姆，S.（1984年）欧洲生物化学学会联合会快报169，194 - 198]以及在Asn334/341处也有发现。N - 乙酰乳糖胺聚糖，主要是部分交叉连接且岩藻糖基化的NeuAcα2→3或Galα1→3Galβ1→4GlcNAcβ1→2Manα1→6(NeuAcα2→6或NeuAcα2→3Gal - β1→4GlcNAcβ1→2Manα1→3)Manβ1→4GlcNacβ1→4GlcNAcβ1→，在6位Manα处有一些分支，是从Asn266、Asn302、Asn334/341、Asn374和Asn410获得的。此外，发现Thr268、Thr277、Thr279、Thr304/309以及Ser273和Ser275被Galβ1→3GalNAcα1→以O - 糖苷键取代，在Gal的3位或/和GalNAc的6位单唾液酸化或去唾液酸化。
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