Molecular Spectroscopic Study on the Interaction between Heparin and Neutral Red

The interaction between heparin and neutral red was investigated by molecular spectroscopic methods. The change of all spectra suggested that positively charged neutral red had interacted with negatively charged heparin. The study of influence factors indicated that electrostatic force and hydrophobic bond might be involved in the interaction. The total binding number per disaccharide unit and intrinsic binding constant were obtained using Scatchard model.     

14C-寡糖在西瓜幼苗植株体内吸收传导和分布

应用同位素示踪技术研究了14C-寡糖在西瓜幼苗植株体内的吸收、传导和分布行为.自显影结果显示,寡糖通过处理叶部或根部后能够被西瓜幼苗植株快速吸收,在叶片中的传导表现为从叶缘向叶片中心分布的趋势.将叶部处理8h和根部处理24h后,14C-寡糖即可以传导和分布到西瓜幼苗的整个植株体内,证明14C-寡糖在西瓜幼苗植株体内具有较强的扩散和向基或向顶传导特征.结果表明,处理叶部4～120h时,根系、茎与未被直接处理的叶片等其它部位的放射性比活度分别由0.18×105和23.08×105Bq/kg变化为0.32×105和3.02×105Bq/kg,总体上表现出向基传导和分布的态势.处理根部4～120h时,西瓜幼苗植株根系、茎部、子叶和真叶中放射性比活度分别由22.23×105,2.23×105,8.33×105和12.78×105Bq/kg变化为431.11×105,42.23×105,65.57×105和78.89×105Bq/kg,表现出14C-寡糖在西瓜幼苗植株体内向顶传导作用和在地上部的积累态势很强.     

Amino acid end-group in commercial heparin. III. Determination of the number of amino groups of amino acid and hexosamine residues per heparin molecule

The reactions of heparin with 2,4,6-trinitrobenzenesulfonic acid (TNBS) were studied spectrometrically. Seven different commercial heparins were used in this study. The amino groups react with TNBS to form equimolar amounts of trinitrophenylated (TNP) amino groups and bisulfite ions. The TNP-amino groups further react with bisulfite ions to form the monosubstituted anionic sigma complex. The absorption spectrum with two maxima at approximately 350 nm and approximately 420 nm, characteristic of either the TNP-amino groups or the complex, was analyzed for the reaction of TNBS with heparin. It was shown that the reactivities of TNBS with amino groups from α-amino acid and hexosamine residues are greatly different. By combining the results of the reaction kinetics and the reaction of heparin with Sanger's reagent, the number of the α-amino groups and the free amino groups in hexosamine residues were determined. These data have been performed with a range of heparins from different commercial sources, of different activities and physical characteristics. No correlation was found between the free amino contents of these heparins and biological potency. © 1993 John Wiley & Sons, Inc.     

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry

A capillary electrophoresis (CE) method was developed for the separation of heparin oligosaccharides compatible to study the interactions between the oligosaccharides and granulocyte-colony stimulating factor (G-CSF). Unfractionated heparin was eliminitively degraded to heparin oligosaccharides by an endolytic heparinase. The degraded smaller oligosaccharides (M(r) < 1000) were baseline-separated by CE under a 50 mM phosphate buffer (pH 9.0) in 10 min. Standard heparin disaccharides and larger oligosaccharides (1000 < M(r) < 8000) were all separated under optimized separation conditions. Compared with standard heparin disaccharides, smaller oligosaccharides contained one nonsulfated, two monosulfated, and two disulfated disaccharides, but trisulfated disaccharides were not found. The smaller oligosaccharides were also identified and molecular mass was deduced by electrospray ionization-mass spectrometry (ESI-MS). Furthermore, interactions between G-CSF and the oligosaccharides were studied by using capillary zone electrophoresis (CZE) under the above separation conditions. It was found that larger oligosaccharides could interact with G-CSF while smaller oligosaccharides were not observed to bind to G-CSF under the experimental conditions. In conclusion, the purified heparinase could selectively degrade heparin into oligosaccharides and the interaction between G-CSF and heparin was correlated with the chain length of heparin.     

An improvement in the bending ability of a hinged trisaccharide with the assistance of a sugar-sugar interaction

Hinged di- and trisaccharides incorporating 2,4-diamino-beta-D-xylopyranoside as a hinge unit (Hin) were synthesized. Bridging of the diamino group of Hin by carbonylation or chelation to a metal ion results in a conformational change from (4)C1 to (1)C4, which in turn causes a bending of the oligosaccharides. In this study, the bending abilities of the hinged oligosaccharides were compared, in terms of the reactivities toward carbonylation and chelation. Di- or trisaccharides containing a 6-O-glycosylated mannopyranoside or galactopyranoside at their reducing ends had bending abilities similar to that of the Hin monosaccharide, probably because there were neither attractive nor repulsive interactions between the reducing and nonreducing ends. However, when Hin was attached at O2 of methyl mannopyranoside (Man alphaMe), the bending ability was dependent on the nonreducing sugar and the reaction conditions. Typically, a disaccharide--Hin beta(1,2)Man alphaMe--was difficult to bend under all the tested reaction conditions, and the bent population in the presence of Zn(II) was only 4%. On the other hand, a trisaccharide--Man alpha(1,3)Hin beta(1,2)Man alphaMe--was bent immediately after the addition of Zn(II) or Hg(II), and the bent population reached 75%, much larger than those of all the other hinged trisaccharides ever tested (<40%). This excellent bending ability suggests an attractive interaction between the reducing and nonreducing ends. The extended conformation was recovered by the addition of triethylenetetramine, a metal ion chelator. Reversible, quick, and efficient bending of the hinged trisaccharide was thus achieved.     

Triazole: the Keystone in Glycosylated Molecular Architectures Constructed by a Click Reaction

The copper(I)‐catalyzed modern version of the Huisgen‐type azide–alkyne cycloaddition to give a 1,4‐disubstituted 1,2,3‐triazole unit is introduced as a powerful ligation method for glycoconjugation. Owing to its high chemoselectivity and tolerance of a variety of reaction conditions, this highly atom‐economic and efficient coupling reaction is especially useful for the effective construction of complex glycosylated structures such as clusters, dendrimers, polymers, peptides, and macrocycles. In all cases the triazole ring plays a key role by locking into position the various parts of these molecular architectures. The examples reported and briefly discussed in this short review highlight the use of this reaction in carbohydrate chemistry and pave the way to further developments and applications.     

Parallel synthesis of multi-branched oligosaccharides related to elicitor active pentasaccharide in rice cell based on orthogonal deprotection and glycosylation strategy

We describe a parallel and efficient synthesis of multi-branched oligosaccharides 3a-g based upon the structure of the phytoalexin elicitor active branched pentasaccharide 2. One-pot sequential orthogonal deprotection of tetrasaccharide 5 with three different protecting groups provided each of seven glycosyl acceptors 4a-g. Glycosylation of the acceptors 4a-g, followed by deprotection provided branched oligosaccharides 3a-g. All the reaction processes from scaffold 5 to 3a-g except for final hydrogenolysis were achieved utilizing an automated synthesizer in a parallel fashion.     

Assembly of a series of malarial glycosylphosphatidylinositol anchor oligosaccharides

We report an efficient and convergent synthesis of a series of oligosaccharides comprised of the malaria GPI glycan (2a), a promising anti-malaria vaccine candidate currently in preclinical trials and several related oligosaccharide sequences (3-8) that are possible biosynthetic precursors of the malarial GPI. A flexible synthetic strategy is disclosed that relies on a late-stage coupling between oligomannosides of varying length and pseudo-disaccharide glycosyl acceptor 11 to readily access various malarial GPI structures. Phosphorylation was accomplished by mild and efficient H-phosphonate chemistry before the final deprotection was carried out by using sodium in ammonia. The direct connection of a thiol group via a phosphate diester linkage to the inositol moiety provides a handle for easy conjugation of the GPI glycan to carrier proteins, immobilization on carbohydrate microarrays and photo-affinity labels identification. These synthetic oligosaccharides will serve as molecular probes.     

Total Synthesis of Sialylated Glycans Related to Avian and Human Influenza Virus Infection

Human and avian influenza type A viruses bind sialylated pentasaccharides. Herein, the total synthesis of four of these glycans is reported. Efficient sialylations relied on two N‐Troc‐protected (Troc=2,2,2‐trichloroethoxycarbonyl) sialic acid building blocks. The first, a thiophenyl glycoside, readily produced the sialyl‐α(2‐6)galactose disaccharide. Combination of the second building block, a novel glycosyl phosphite, and a benzylidene‐protected galactoside produced the best results for the formation of the sialyl‐α(2‐3)galactose. Two common trisaccharides were assembled by the introduction of glucose, galactose, and glucosamine building blocks followed by selective deprotection. Two sets of pentasaccharides were obtained by the union of two sialylgalactose N‐phenyl trifluoroacetimidate building blocks with the two trisaccharides above. Global deprotection furnished the desired pentasaccharides. The products of these total syntheses are currently employed on the surface of carbohydrate microarrays to detect and type different strains of the influenza virus.