首页> 美国卫生研究院文献>Journal of Bacteriology >Structural studies of lipooligosaccharides from Haemophilus ducreyi ITM 5535 ITM 3147 and a fresh clinical isolate ACY1: evidence for intrastrain heterogeneity with the production of mutually exclusive sialylated or elongated glycoforms.
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Structural studies of lipooligosaccharides from Haemophilus ducreyi ITM 5535 ITM 3147 and a fresh clinical isolate ACY1: evidence for intrastrain heterogeneity with the production of mutually exclusive sialylated or elongated glycoforms.

机译:来自杜克嗜血杆菌ITM 5535ITM 3147和新鲜的临床分离株ACY1的脂寡糖的结构研究:菌株内异质性的证据包括相互排斥的唾液酸化或细长糖型的产生。

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摘要

The structures of the lipooligosaccharides (LOSs) from Haemophilus ducreyi ITM 5535 and ITM 3147 and a fresh clinical isolate, ACY1, have been investigated. Oligosaccharides were obtained from phenol-water-extracted LOS by mild acid hydrolysis and were studied by methylation analysis, fast atom bombardment and electrospray ionization mass spectrometry, and nuclear magnetic resonance spectroscopy. The major oligosaccharide obtained from all strains was a nonasaccharide with the structure beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-D-Galp-(1-->4)-D-a lpha-D-Hepp- (1-->6)-beta-D-Glcp-(1-->[L-alpha-D-Hepp-(1-->2)-L-alpha-D-Hepp - (1-->3)]4)-L-alpha-D-Hepp-Kdo (Kdo stands for 3-deoxy-D-manno-octulosonic acid) and is thus identical to that identified as the major oligosaccharide in H. ducreyi ITM 2665 (E. K. H. Schweda, A. C. Sundström, L. M. Eriksson, J.A. Jonasson, and A. A. Lindberg, J. Biol. Chem. 269:12040-12048, 1994). Electrospray ionization mass spectrometry on O-deacylated LOS from H. ducreyi ITM 5535 obtained after treatment with anhydrous hydrazine gave evidence for the presence of a sialylated major compound, Neu5Ac alpha(2-->3)-beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-D-Gal p- (1-->4)-D-alpha-D-Hepp-(1-->6)-beta-D-Glcp-(1-->[L-alpha-D-Hepp -(1-->2)-L- alpha-D-Hepp-(1-->3)]4)-L-alpha-D-Hepp-Kdo(P)-O-deacylated lipid A (Neu5Ac stands for N-acetylneuraminic acid). However, an even larger oligosaccharide could be isolated from all strains as a minor component, viz., the undecasaccharide beta-D-Galp-(1-->4)-beta-D-GlcNAcp-(1-->3)-beta-d-Galp-(1-->4)-beta-D-glcNAcp-(1-->3)-beta-D-Galp-(1-->4)-D-alpha-D-Hepp-(1-->6)-beta-D-Glcp-(1-->[L-alpha-D-Hepp-(1-->2)-L-alpha-D-Hepp-(1-->3)]4-L-alpha-D-Hepp-Kdo, which represents an N-acetyl lactosamine disaccharide unit elongation of the LOS outer core. No Sialylation of this latter minor component undecasaccharide was detected.
机译:研究了杜克嗜血杆菌ITM 5535和ITM 3147的脂寡糖(LOS)以及新鲜的临床分离株ACY1的结构。寡糖是通过苯酚-水萃取的LOS通过轻度酸水解而获得的,并通过甲基化分析,快速原子轰击和电喷雾电离质谱法以及核磁共振波谱进行了研究。从所有菌株获得的主要寡糖是结构为β-D-Galp-(1-> 4)-beta-D-GlcNAcp-(1-> 3)-beta-D-Galp-(1-的九糖-> 4)-Da lpha-D-Hepp-(1-> 6)-beta-D-Glcp-(1-> [L-alpha-D-Hepp-(1-> 2)-L- alpha-D-Hepp-(1-> 3)] 4)-L-alpha-D-Hepp-Kdo(Kdo代表3-deoxy-D-manno-octulosonic acid),因此与确定为H. ducreyi ITM 2665中的主要低聚糖(EKH Schweda,ACSundström,LM Eriksson,JA Jonasson,and AA Lindberg,J.Biol.Chem.269:12040-12048,1994)。用无水肼处理后获得的来自杜克氏杆菌ITM 5535的O-去酰化LOS的电喷雾电离质谱分析表明存在唾液酸化的主要化合物Neu5Acα(2-> 3)-β-D-Galp-(1 -> 4)-beta-D-GlcNAcp-(1-> 3)-beta-D-Gal p-(1-> 4)-D-alpha-D-Hepp-(1-> 6) -beta-D-Glcp-(1-> [L-alpha-D-Hepp-(1-> 2)-L- alpha-D-Hepp-(1-> 3)] 4)-L- α-D-Hepp-Kdo(P)-O-去酰基化脂质A(Neu5Ac代表N-乙酰神经氨酸)。但是,可以从所有菌株中分离出更大的寡糖作为次要成分,即十一糖β-D-Galp-(1-> 4)-β-D-GlcNAcp-(1-> 3)- beta-d-Galp-(1-> 4)-beta-D-glcNAcp-(1-> 3)-beta-D-Galp-(1-> 4)-D-alpha-D-Hepp- (1-> 6)-beta-D-Glcp-(1-> [L-alpha-D-Hepp-(1-> 2)-L-alpha-D-Hepp-(1-> 3 )]4-L-α-D-Hepp-Kdo,代表LOS外核的N-乙酰基乳糖胺二糖单元延伸,未检测到后者次要成分十一碳糖的唾液酸化。

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