Pub Date : 2009-03-18DOI: 10.2174/1875398100902010016
R. Miranda
The use of natural gums, taken from the exudates and extracts of plants, has been given a strong impulse due to both the many and lucrative possibilities for industrialization and to the excellent international market. A gum or resin of a yellowish color, soluble in water, and which presents a great potential for industrialization, appears on the trunk and branches of the cashew tree; that is known mainly for its nuts that are used as a food ingredient. Cashew gum interacts with water and it has emulsifier, adhesive and stabilizer properties, occurring in the form of pale yellow to reddish am- bers. In cold water, it swells into a jelly like mass but dissolves rapidly when heated. The resin/gum secretory ducts that appear along the cashew tree bark were studied using histological procedures and microscopic observations. The gum was studied at its crude state and then chemically treated in order to obtain a kind of gel (microbiofilm).
{"title":"Cashew Tree Bark Secretion - Persectives for its Use in Protein Isolation Strategies","authors":"R. Miranda","doi":"10.2174/1875398100902010016","DOIUrl":"https://doi.org/10.2174/1875398100902010016","url":null,"abstract":"The use of natural gums, taken from the exudates and extracts of plants, has been given a strong impulse due to both the many and lucrative possibilities for industrialization and to the excellent international market. A gum or resin of a yellowish color, soluble in water, and which presents a great potential for industrialization, appears on the trunk and branches of the cashew tree; that is known mainly for its nuts that are used as a food ingredient. Cashew gum interacts with water and it has emulsifier, adhesive and stabilizer properties, occurring in the form of pale yellow to reddish am- bers. In cold water, it swells into a jelly like mass but dissolves rapidly when heated. The resin/gum secretory ducts that appear along the cashew tree bark were studied using histological procedures and microscopic observations. The gum was studied at its crude state and then chemically treated in order to obtain a kind of gel (microbiofilm).","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"2 1","pages":"16-19"},"PeriodicalIF":0.0,"publicationDate":"2009-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-01-29DOI: 10.2174/1875398100902010001
Z. Rasheed, L. kumar, I. Prasad, Nadeem Ansari
The role of advanced glycation end products (AGEs)-damaged immunoglobulin G (AGE-IgG) in type 1 diabe- tes has been investigated in the present study. IgG was isolated from the normal humans and was subjected to in vitro gly- cation with glucose. The AGEs caused extensive damaged to IgG. The AGE-IgG was found to be highly immunogenic in rabbits as compared to native IgG. The binding characteristics of circulating autoantibodies in type 1 diabetes mellitus (DM) patients against native and AGE-IgG were assessed. Type 1 DM patients (n=31) were examined by ELISA and their results were compared with healthy age-matched human controls (n=22). High degree of specific binding by 61.3 % of DM sera autoantibodies towards AGE-IgG was observed, in comparison to its native analog (p< 0.05). Sera from those type 1 DM patients having smoking history, high aging with high degree of disease showed substantially stronger binding to AGE-IgG over native IgG in particular. IgG from type 1 DM patients (DM-IgG) contained higher levels of carbonyls as compared to normal human subjects (normal-IgG) (p<0.001). Collectively, the AGEs modification of IgG causes pertur- bations, resulting in the generation of neo-epitopes, and making it a potential immunogen. The IgG modified with AGEs may be one of the factors for the induction of circulating type 1 diabetes autoantibodies.
{"title":"Advanced Glycation End Products (AGEs) Damaged IgG, a Target for Circulating Autoantibodies in Patients with Type 1 Diabetes Mellitus","authors":"Z. Rasheed, L. kumar, I. Prasad, Nadeem Ansari","doi":"10.2174/1875398100902010001","DOIUrl":"https://doi.org/10.2174/1875398100902010001","url":null,"abstract":"The role of advanced glycation end products (AGEs)-damaged immunoglobulin G (AGE-IgG) in type 1 diabe- tes has been investigated in the present study. IgG was isolated from the normal humans and was subjected to in vitro gly- cation with glucose. The AGEs caused extensive damaged to IgG. The AGE-IgG was found to be highly immunogenic in rabbits as compared to native IgG. The binding characteristics of circulating autoantibodies in type 1 diabetes mellitus (DM) patients against native and AGE-IgG were assessed. Type 1 DM patients (n=31) were examined by ELISA and their results were compared with healthy age-matched human controls (n=22). High degree of specific binding by 61.3 % of DM sera autoantibodies towards AGE-IgG was observed, in comparison to its native analog (p< 0.05). Sera from those type 1 DM patients having smoking history, high aging with high degree of disease showed substantially stronger binding to AGE-IgG over native IgG in particular. IgG from type 1 DM patients (DM-IgG) contained higher levels of carbonyls as compared to normal human subjects (normal-IgG) (p<0.001). Collectively, the AGEs modification of IgG causes pertur- bations, resulting in the generation of neo-epitopes, and making it a potential immunogen. The IgG modified with AGEs may be one of the factors for the induction of circulating type 1 diabetes autoantibodies.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"2 1","pages":"1-8"},"PeriodicalIF":0.0,"publicationDate":"2009-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-01-29DOI: 10.2174/1875398100902010009
R. Ward
A method was developed to isolate mono- and disaccharide-free oligosaccharides from human and bovine milk using a combination of enzymatic digestion of lactose and solid-phase extraction. In the initial trial, 2.5 g of oligosaccha- rides were isolated from one liter of human milk. In subsequent trials this was increased to over 5 g of oligosaccharides per liter. Compared to filtration-based extraction methods, this procedure allows for further isolation of oligosaccharide fractions via modulation of the column rinsing solvent. Neutral monosaccharide composition of the oligosaccharide poly- mers was investigated using gas chromatographic analysis of the monosaccharides as alditol acetate derivatives. Results indicate oligosaccharides are approximately made up of 24% fucose, 41% galactose, 22% glucose and 13% glucosamine. Isolated bovine and human milk oligosaccharides were compared to lactose as fermentation substrates for Bifidobacterium longum biovar infantis. Lactose fermentation yielded the greatest production of biomass followed by bovine and human milk oligosaccharides.
{"title":"Isolation of Milk Oligosaccharides using Solid-Phase Extraction","authors":"R. Ward","doi":"10.2174/1875398100902010009","DOIUrl":"https://doi.org/10.2174/1875398100902010009","url":null,"abstract":"A method was developed to isolate mono- and disaccharide-free oligosaccharides from human and bovine milk using a combination of enzymatic digestion of lactose and solid-phase extraction. In the initial trial, 2.5 g of oligosaccha- rides were isolated from one liter of human milk. In subsequent trials this was increased to over 5 g of oligosaccharides per liter. Compared to filtration-based extraction methods, this procedure allows for further isolation of oligosaccharide fractions via modulation of the column rinsing solvent. Neutral monosaccharide composition of the oligosaccharide poly- mers was investigated using gas chromatographic analysis of the monosaccharides as alditol acetate derivatives. Results indicate oligosaccharides are approximately made up of 24% fucose, 41% galactose, 22% glucose and 13% glucosamine. Isolated bovine and human milk oligosaccharides were compared to lactose as fermentation substrates for Bifidobacterium longum biovar infantis. Lactose fermentation yielded the greatest production of biomass followed by bovine and human milk oligosaccharides.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"2 1","pages":"9-15"},"PeriodicalIF":0.0,"publicationDate":"2009-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-12-16DOI: 10.2174/1875398100801010052
Arivalagan Muthusamy, D. Gowda, V. P. Bhavanandan
Epithelial mucin glycoproteins of bladder act as an effective barrier against invasion by pathogenic microor- ganisms and injury by toxic substances in urine. Although these glycoconjugates play important roles in the pathophysiol- ogy of bladder disorders such as intestinal cystisis, cancer, and urinary tract infections, they have not been characterized in detail either in humans or in animals. Rabbits could be useful for developing models for studying bladder disorders. In this study, we purified and partially characterized two major high molecular weight rabbit bladder mucin glycoproteins, desig- nated RBM1 and RBM2, found in urine. Consistent with their mucin characteristics, amino acid compositions showed have high levels of serine, glutamic acid, proline, glycine and alanine, which together comprise 34% and 42% of the total amino acids in RBM1 and RBM2, respectively. Carbohydrate compositional analysis indicated that RBM1 and RBM2 con- sist of N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), galactose (Gal), N-acetylneuraminic acid (NeuAc) and fucose (Fuc) in the molar ratio of 1.0: 0.82: 0.12: 0.30: 0.02 and 1.0: 1.03: 0.46: 0.16: 0.05, respectively; mannose (Man) was not detected in either mucin. Both mucin fractions were strongly reactive to wheat germ agglutinin, but not to Ca2 antibody specific to a human tumor mucin antigen (asialylated carbohydrate linked to protein core), sug- gesting that most of the galactosyl residues of oligosaccharides are sialylated. Together, the data suggest that rabbit mucin glycoproteins characterized here are distinctively different from MUC1 mucin glycoprotein found in human urine.
{"title":"Characterization of High Molecular Weight Mucins of Rabbit Bladder","authors":"Arivalagan Muthusamy, D. Gowda, V. P. Bhavanandan","doi":"10.2174/1875398100801010052","DOIUrl":"https://doi.org/10.2174/1875398100801010052","url":null,"abstract":"Epithelial mucin glycoproteins of bladder act as an effective barrier against invasion by pathogenic microor- ganisms and injury by toxic substances in urine. Although these glycoconjugates play important roles in the pathophysiol- ogy of bladder disorders such as intestinal cystisis, cancer, and urinary tract infections, they have not been characterized in detail either in humans or in animals. Rabbits could be useful for developing models for studying bladder disorders. In this study, we purified and partially characterized two major high molecular weight rabbit bladder mucin glycoproteins, desig- nated RBM1 and RBM2, found in urine. Consistent with their mucin characteristics, amino acid compositions showed have high levels of serine, glutamic acid, proline, glycine and alanine, which together comprise 34% and 42% of the total amino acids in RBM1 and RBM2, respectively. Carbohydrate compositional analysis indicated that RBM1 and RBM2 con- sist of N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), galactose (Gal), N-acetylneuraminic acid (NeuAc) and fucose (Fuc) in the molar ratio of 1.0: 0.82: 0.12: 0.30: 0.02 and 1.0: 1.03: 0.46: 0.16: 0.05, respectively; mannose (Man) was not detected in either mucin. Both mucin fractions were strongly reactive to wheat germ agglutinin, but not to Ca2 antibody specific to a human tumor mucin antigen (asialylated carbohydrate linked to protein core), sug- gesting that most of the galactosyl residues of oligosaccharides are sialylated. Together, the data suggest that rabbit mucin glycoproteins characterized here are distinctively different from MUC1 mucin glycoprotein found in human urine.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"1 1","pages":"52-57"},"PeriodicalIF":0.0,"publicationDate":"2008-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-10-15DOI: 10.2174/1875398100801010040
T. Schürpf, N. Callewaert, M. Meyer, C. Tränkle, W. Laroy, R. Cummings, V. Otto
Soluble intercellular adhesion molecule-1 (sICAM-1) is elevated in the cerebrospinal fluid of patients with se- vere brain trauma and mouse sICAM-1 induces the production of macrophage inflammatory protein-2 (MIP-2) in mouse astrocytes. The production of MIP-2 is greatly enhanced when sICAM-1 contains sialylated complex-type N-glycans (sICAM-1-CT) as produced by Chinese hamster ovary (CHO) cells. By contrast, sICAM-1 from the Lec1 mutant of CHO cells (sICAM-1-HM), containing only high mannose-type N-glycans, is relatively inactive. Here we show that the N- glycans of sICAM-1-CT are mostly 2,3-sialylated bi-, tri-, and tetraantennary complex-type structures with varying amounts of core fucosylation. Unexpectedly, sICAM-1-CT and sICAM-1-HM bound equivalently to mouse astrocytes. Enhanced MIP-2 induction by sICAM-1-CT was associated with a more rapid, higher level, and prolonged MIP-2 re- sponse as well as sICAM-1-CT accumulation at the plasma membranes of mouse astrocytes. These results show that gly- cosylation of sICAM-1 contributes to its signaling properties at the astrocyte cell surface, and suggest that altered glyco- sylation which might arise as a result of inflammation could regulate the bioactivity of sICAM-1.
{"title":"Consequences of soluble ICAM-1 N-glycan alterations on receptor binding and signaling kinetics in mouse astrocytes","authors":"T. Schürpf, N. Callewaert, M. Meyer, C. Tränkle, W. Laroy, R. Cummings, V. Otto","doi":"10.2174/1875398100801010040","DOIUrl":"https://doi.org/10.2174/1875398100801010040","url":null,"abstract":"Soluble intercellular adhesion molecule-1 (sICAM-1) is elevated in the cerebrospinal fluid of patients with se- vere brain trauma and mouse sICAM-1 induces the production of macrophage inflammatory protein-2 (MIP-2) in mouse astrocytes. The production of MIP-2 is greatly enhanced when sICAM-1 contains sialylated complex-type N-glycans (sICAM-1-CT) as produced by Chinese hamster ovary (CHO) cells. By contrast, sICAM-1 from the Lec1 mutant of CHO cells (sICAM-1-HM), containing only high mannose-type N-glycans, is relatively inactive. Here we show that the N- glycans of sICAM-1-CT are mostly 2,3-sialylated bi-, tri-, and tetraantennary complex-type structures with varying amounts of core fucosylation. Unexpectedly, sICAM-1-CT and sICAM-1-HM bound equivalently to mouse astrocytes. Enhanced MIP-2 induction by sICAM-1-CT was associated with a more rapid, higher level, and prolonged MIP-2 re- sponse as well as sICAM-1-CT accumulation at the plasma membranes of mouse astrocytes. These results show that gly- cosylation of sICAM-1 contributes to its signaling properties at the astrocyte cell surface, and suggest that altered glyco- sylation which might arise as a result of inflammation could regulate the bioactivity of sICAM-1.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"1 1","pages":"40-51"},"PeriodicalIF":0.0,"publicationDate":"2008-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-06-06DOI: 10.2174/1875398100801010025
N. Kawazoe, H. Okada, E. Fukushi, A. Yamamori, S. Onodera, J. Kawabata, N. Shiomi
Fermented beverage of plant extract was prepared from about fifty kinds of vegetables and fruits. Natural fermentation was carried out mainly by lactic acid bacteria (Leuconostoc spp.) and yeast (Zygosaccharomyces spp.and Pichia spp.). Two novel oligosaccharides have been found from this beverage and isolated from the beverage using carbon-Celite column chromatography and preparative high performance liquid chromatography. Structure confirmation of the saccharides was provided by methylation analysis, MALDI-TOF-MS and NMR measurements. These saccharides were identified as new trisaccharides, -D-glucopyranosyl-(1 1)-D-fructofuranosyl-(2 1)-D-glucopyranoside; -Dgalactopyranosyl-(1 1)-D-fructofuranosyl-(2 1)-D-glucopyranoside.
以50余种蔬菜和水果为原料，制备了植物提取物发酵饮料。自然发酵主要由乳酸菌(Leuconostoc spp.)和酵母(Zygosaccharomyces spp.和Pichia spp.)进行。从该饮料中发现了两种新的低聚糖，并采用碳柱色谱法和制备型高效液相色谱法从该饮料中分离得到。通过甲基化分析、MALDI-TOF-MS和核磁共振测量证实了糖的结构。这些糖被鉴定为新的三糖:- d -葡萄糖吡喃糖基-(11)- d -呋喃糖基-(21)- d -葡萄糖吡喃糖苷;- dgalactopyranosyl -(11)- d -fructofuranosyl-(21)- d -glucopyranoside
{"title":"Structural Analysis of Two Trisaccharides Isolated from Fermented Beverage of Plant Extract","authors":"N. Kawazoe, H. Okada, E. Fukushi, A. Yamamori, S. Onodera, J. Kawabata, N. Shiomi","doi":"10.2174/1875398100801010025","DOIUrl":"https://doi.org/10.2174/1875398100801010025","url":null,"abstract":"Fermented beverage of plant extract was prepared from about fifty kinds of vegetables and fruits. Natural fermentation was carried out mainly by lactic acid bacteria (Leuconostoc spp.) and yeast (Zygosaccharomyces spp.and Pichia spp.). Two novel oligosaccharides have been found from this beverage and isolated from the beverage using carbon-Celite column chromatography and preparative high performance liquid chromatography. Structure confirmation of the saccharides was provided by methylation analysis, MALDI-TOF-MS and NMR measurements. These saccharides were identified as new trisaccharides, -D-glucopyranosyl-(1 1)-D-fructofuranosyl-(2 1)-D-glucopyranoside; -Dgalactopyranosyl-(1 1)-D-fructofuranosyl-(2 1)-D-glucopyranoside.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"1 1","pages":"25-30"},"PeriodicalIF":0.0,"publicationDate":"2008-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-05-30DOI: 10.2174/1875398100801010019
F. Jamoisa, F. Goffic, J. Yvin, D. Plusquellec, V. Ferrières
Laminaribiose, which is the simplest -(1,3)-glucan, is one of the most powerful agents able to increase germi- nation. Its chemical synthesis was revised in detail starting from peracylated donors and easily available glucofuranose protected by acetal groups in the presence of appropriated catalyst and/or promoter. We particularly focused our attention on the nature of the Lewis acid generally required in glycosidic couplings. Finally, an interesting scale-up was performed which allowed us to prepare laminaribiose on a kilogram scale.
{"title":"How to Improve Chemical Synthesis of Laminaribiose on a Large Scale","authors":"F. Jamoisa, F. Goffic, J. Yvin, D. Plusquellec, V. Ferrières","doi":"10.2174/1875398100801010019","DOIUrl":"https://doi.org/10.2174/1875398100801010019","url":null,"abstract":"Laminaribiose, which is the simplest -(1,3)-glucan, is one of the most powerful agents able to increase germi- nation. Its chemical synthesis was revised in detail starting from peracylated donors and easily available glucofuranose protected by acetal groups in the presence of appropriated catalyst and/or promoter. We particularly focused our attention on the nature of the Lewis acid generally required in glycosidic couplings. Finally, an interesting scale-up was performed which allowed us to prepare laminaribiose on a kilogram scale.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"1 1","pages":"19-24"},"PeriodicalIF":0.0,"publicationDate":"2008-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-05-07DOI: 10.2174/1875398100801010008
H. Yamada, Erina Ohno, M. Utsumi, Y. Yamaguchi, E. Kurimoto, N. Takahashi, S. Oka, T. Kawasaki, Koichi Kato
Although the multi-dimensional HPLC maps of neutral, sialyl, and sulfated N-glycans have been reported and widely used for glycosylation profiling, those of glucuronyl oligosaccharides have not yet been available. In the present study, by in vitro enzymatic reactions, we prepared 55 different glucuronyl PA-oligosaccharides that include 6 kinds of HNK-1-containing N-glycans, and established their HPLC map. Furthermore, we applied this map to the characterization of branch specificity in glucuronylation reaction catalyzed by human GlcAT-S, revealing that this enzyme transfers the glucuronyl residues preferentially onto the Gal 1�4GlcNAc 1�4Man 1�3 and Gal 1�4GlcNAc 1�2Man 1�3 branches of a galactose-terminated tri-antennary oligosaccharide. The HPLC map developed in the present study will be a useful glycomics tool for identification and profiling of glucuronyl N-glycans expressed in the neural and other biological systems.
{"title":"Development and Application of High Performance Liquid Chromatography Map of Glucuronyl N-glycans","authors":"H. Yamada, Erina Ohno, M. Utsumi, Y. Yamaguchi, E. Kurimoto, N. Takahashi, S. Oka, T. Kawasaki, Koichi Kato","doi":"10.2174/1875398100801010008","DOIUrl":"https://doi.org/10.2174/1875398100801010008","url":null,"abstract":"Although the multi-dimensional HPLC maps of neutral, sialyl, and sulfated N-glycans have been reported and widely used for glycosylation profiling, those of glucuronyl oligosaccharides have not yet been available. In the present study, by in vitro enzymatic reactions, we prepared 55 different glucuronyl PA-oligosaccharides that include 6 kinds of HNK-1-containing N-glycans, and established their HPLC map. Furthermore, we applied this map to the characterization of branch specificity in glucuronylation reaction catalyzed by human GlcAT-S, revealing that this enzyme transfers the glucuronyl residues preferentially onto the Gal 1�4GlcNAc 1�4Man 1�3 and Gal 1�4GlcNAc 1�2Man 1�3 branches of a galactose-terminated tri-antennary oligosaccharide. The HPLC map developed in the present study will be a useful glycomics tool for identification and profiling of glucuronyl N-glycans expressed in the neural and other biological systems.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"1 1","pages":"8-18"},"PeriodicalIF":0.0,"publicationDate":"2008-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-04-04DOI: 10.2174/1875398100801010001
O. Renaudet, P. Dumy
The fully solid-phase synthesis of chemically well-defined glycoclusters grafted to a topological cyclodecapep- tide template is described. The orthogonally protected peptide backbone was first synthesized and cyclized on solid sup- port using D-glutamic acid as first amino acid linked to the resin. After successive regioselective deprotection steps, bi- otins were coupled to the lower addressable domains of the scaffold, then carbohydrates-binding ligands were assembled as cluster on the upper domain using a chemoselective oxime-based strategy. This provides multitopic labeled glycopep- tides which can be easily immobilized to streptavidin-coated surfaces for studying carbohydrate-protein interactions in glycomic researches.
{"title":"A Fully Solid-Phase Synthesis of Biotinylated Glycoclusters","authors":"O. Renaudet, P. Dumy","doi":"10.2174/1875398100801010001","DOIUrl":"https://doi.org/10.2174/1875398100801010001","url":null,"abstract":"The fully solid-phase synthesis of chemically well-defined glycoclusters grafted to a topological cyclodecapep- tide template is described. The orthogonally protected peptide backbone was first synthesized and cyclized on solid sup- port using D-glutamic acid as first amino acid linked to the resin. After successive regioselective deprotection steps, bi- otins were coupled to the lower addressable domains of the scaffold, then carbohydrates-binding ligands were assembled as cluster on the upper domain using a chemoselective oxime-based strategy. This provides multitopic labeled glycopep- tides which can be easily immobilized to streptavidin-coated surfaces for studying carbohydrate-protein interactions in glycomic researches.","PeriodicalId":88944,"journal":{"name":"Open glycoscience","volume":"1 1","pages":"1-7"},"PeriodicalIF":0.0,"publicationDate":"2008-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68111077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-01-01DOI: 10.2174/1875398100801010031
Sarah B Frazier, Kevin A Roodhouse, Dennis E Hourcade, Lijuan Zhang
Glycosaminoglycans (GAGs) are linear polysaccharides that are found in the extracellular matrix and biological fluids of animals where they interact with hundreds of proteins and perform a variety of critical roles. There are five classes of animal GAGs: heparan sulfate (HS), chondroitin sulfate (CS), dermatan sulfate (DS), keratan sulfate (KS), and hyaluronan (HA). Many biological functions can be monitored directly by their impact on GAG quantity. Thus, simple, sensitive, and robust GAG quantification methods are needed for the development of biomarkers. We have systematically compared three available GAG quantification assays including an HPLC-based assay, a simplified Alcian Blue assay, and a miniaturized carbazole assay. The carbazole and Alcian Blue assays were reproducible and simple to perform in general lab settings, but had important limitations: The carbazole assay could not detect KS and it overestimated GAGs that were contaminated with salts or dissolved in PBS. The Alcian Blue assay detected only those GAGs that were sulfated. In contrast, while the HPLC method was time-consuming, it was a robust and sensitive assay that not only detected all GAGs but also quantified glucosamine-GAGs and galactosamine-GAGs simultaneously. The HPLC assay was not affected by salt or level of GAG sulfation and it yielded reproducible values for all types of GAGs tested. These results suggest that an automated HPLC assay would be generally useful for the routine measurement of a panel of GAG-based biomarkers while the carbazole assay and the Alcian Blue assays could prove valuable for more specific purposes.
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