Glycobiology最新文献_第10页

Reusable glycan microarrays using a microwave assisted wet-erase (MAWE) process. 使用微波辅助湿擦(MAWE)工艺的可重复使用聚糖微阵列。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-26 DOI: 10.1093/glycob/cwad091

Akul Y Mehta, Catherine A Tilton, Lukas Muerner, Stephan von Gunten, Jamie Heimburg-Molinaro, Richard D Cummings

{"title":"Reusable glycan microarrays using a microwave assisted wet-erase (MAWE) process.","authors":"Akul Y Mehta, Catherine A Tilton, Lukas Muerner, Stephan von Gunten, Jamie Heimburg-Molinaro, Richard D Cummings","doi":"10.1093/glycob/cwad091","DOIUrl":"10.1093/glycob/cwad091","url":null,"abstract":"Modern studies on binding of proteins to glycans commonly involve the use of synthetic glycans and their derivatives in which a small amount of the material is covalently printed onto a functionalized slide in a glycan microarray format. While incredibly useful to explore binding interactions with many types of samples, the common techniques involve drying the slides, which leads to irreversible association of the protein to the spots on slides to which they bound, thus limiting a microarray to a single use. We have developed a new technique which we term Microwave Assisted Wet-Erase (MAWE) glycan microarrays. In this approach we image the slides under wet conditions to acquire the data, after which the slides are cleaned of binding proteins by treatment with a denaturing SDS solution along with microwave treatment. Slides cleaned in this way can be reused multiple times, and an example here shows the reuse of a single array 15 times. We also demonstrate that this method can be used for a single-array per slide or multi-array per slide platforms. Importantly, the results obtained using this technique for a variety of lectins sequentially applied to a single array, are concordant to those obtained via the classical dry approaches on multiple slides. We also demonstrate that MAWE can be used for different types of samples, such as serum for antibody binding, and whole cells, such as yeast. This technique will greatly conserve precious glycans and prolong the use of existing and new glycan microarrays.","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10969520/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"92153571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Developing a solid-phase method for the enzymatic synthesis of heparan sulfate and chondroitin sulfate backbones. 建立固相法酶法合成硫酸肝素和硫酸软骨素骨架。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-26 DOI: 10.1093/glycob/cwad093

Eduardo Stancanelli, Wei Liu, Guowei Su, Vijay Padagala, Jian Liu

引用次数: 0

Glyco-Forum. Glyco-Forum.

IF 4.3 3区生物学

Glycobiology Pub Date : 2024-03-26 DOI: 10.1093/glycob/cwae021

引用次数: 0

Galectin-3 does not interact with RNA directly. 半乳糖凝集素-3不直接与RNA相互作用。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwad076

Egan L Peltan, Nicholas M Riley, Ryan A Flynn, David S Roberts, Carolyn R Bertozzi

{"title":"Galectin-3 does not interact with RNA directly.","authors":"Egan L Peltan, Nicholas M Riley, Ryan A Flynn, David S Roberts, Carolyn R Bertozzi","doi":"10.1093/glycob/cwad076","DOIUrl":"10.1093/glycob/cwad076","url":null,"abstract":"Galectin-3, well characterized as a glycan binding protein, has been identified as a putative RNA binding protein, possibly through participation in pre-mRNA maturation through interactions with splicosomes. Given recent developments with cell surface RNA biology, the putative dual-function nature of galectin-3 evokes a possible non-classical connection between glycobiology and RNA biology. However, with limited functional evidence of a direct RNA interaction, many molecular-level observations rely on affinity reagents and lack appropriate genetic controls. Thus, evidence of a direct interaction remains elusive. We demonstrate that antibodies raised to endogenous human galectin-3 can isolate RNA-protein crosslinks, but this activity remains insensitive to LGALS3 knock-out. Proteomic characterization of anti-galectin-3 IPs revealed enrichment of galectin-3, but high abundance of hnRNPA2B1, an abundant, well-characterized RNA-binding protein with weak homology to the N-terminal domain of galectin-3, in the isolate. Genetic ablation of HNRNPA2B1, but not LGALS3, eliminates the ability of the anti-galectin-3 antibodies to isolate RNA-protein crosslinks, implying either an indirect interaction or cross-reactivity. To address this, we introduced an epitope tag to the endogenous C-terminal locus of LGALS3. Isolation of the tagged galectin-3 failed to reveal any RNA-protein crosslinks. This result suggests that the galectin-3 does not directly interact with RNA and may be misidentified as an RNA-binding protein, at least in HeLa where the putative RNA associations were first identified. We encourage further investigation of this phenomenon employ gene deletions and, when possible, endogenous epitope tags to achieve the specificity required to evaluate potential interactions.","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41198953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Glyco-Forum. Glyco-Forum.

IF 4.3 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwae020

引用次数: 0

N-glycomic profiling of capsid proteins from Adeno-Associated Virus serotypes. 腺相关病毒血清型衣壳蛋白的N-糖蛋白谱分析。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwad074

Yongjing Xie, Michael Butler

{"title":"N-glycomic profiling of capsid proteins from Adeno-Associated Virus serotypes.","authors":"Yongjing Xie, Michael Butler","doi":"10.1093/glycob/cwad074","DOIUrl":"10.1093/glycob/cwad074","url":null,"abstract":"Adeno-associated virus (AAV) vector has become the leading platform for gene delivery. Each serotype exhibits a different tissue tropism, immunogenicity, and in vivo transduction performance. Therefore, selecting the most suitable AAV serotype is critical for efficient gene delivery to target cells or tissues. Genome divergence among different serotypes is due mainly to the hypervariable regions of the AAV capsid proteins. However, the heterogeneity of capsid glycosylation is largely unexplored. In the present study, the N-glycosylation profiles of capsid proteins of AAV serotypes 1 to 9 have been systemically characterized and compared using a previously developed high-throughput and high-sensitivity N-glycan profiling platform. The results showed that all 9 investigated AAV serotypes were glycosylated, with comparable profiles. The most conspicuous feature was the high abundance mannosylated N-glycans, including FM3, M5, M6, M7, M8, and M9, that dominated the chromatograms within a range of 74 to 83%. Another feature was the relatively lower abundance of fucosylated and sialylated N-glycan structures, in the range of 23%-40% and 10%-17%, respectively. However, the exact N-glycan composition differed. These differences may be utilized to identify potential structural relationships between the 9 AAV serotypes. The current research lays the foundation for gaining better understanding of the importance of N-glycans on the AAV capsid surface that may play a significant role in tissue tropism, interaction with cell surface receptors, cellular uptake, and intracellular processing.","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10950483/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41117813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Expression of low-sulfated keratan sulfate in non-mucinous ovarian carcinoma. 非黏液性卵巢癌中低硫酸化角蛋白硫酸盐的表达。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwad056

Hitomi Hoshino, Ya-Ying Chen, Daisuke Inoue, Yoshio Yoshida, Kay-Hooi Khoo, Tomoya O Akama, Motohiro Kobayashi

{"title":"Expression of low-sulfated keratan sulfate in non-mucinous ovarian carcinoma.","authors":"Hitomi Hoshino, Ya-Ying Chen, Daisuke Inoue, Yoshio Yoshida, Kay-Hooi Khoo, Tomoya O Akama, Motohiro Kobayashi","doi":"10.1093/glycob/cwad056","DOIUrl":"10.1093/glycob/cwad056","url":null,"abstract":"Keratan sulfate glycosaminoglycan is composed of repeating N-acetyllactosamine (LacNAc) disaccharide units consisting of galactose (Gal) and N-acetylglucosamine (GlcNAc), both often 6-O-sulfated. Sulfate contents of keratan sulfate are heterogeneous depending upon the origins. In this study, keratan sulfate is classified as either highly sulfated (in which both GlcNAc and Gal residues are 6-O-sulfated) or low-sulfated (in which only GlcNAc residues are 6-O-sulfated). It is reported that highly sulfated keratan sulfate detected by the 5D4 monoclonal antibody is preferentially expressed in normal epithelial cells lining the female genital tract and in their neoplastic counterparts; however, expression of low-sulfated keratan sulfate in either has not been characterized. In the present study, we generated the 294-1B1 monoclonal antibody, which selectively recognizes low-sulfated keratan sulfate, and performed precise glycan analysis of sulfated glycans expressed on human serous ovarian carcinoma OVCAR-3 cells. We found that OVCAR-3 cells do not express highly sulfated keratan sulfate but rather express low-sulfated form, which was heterogeneous in 294-1B1 reactivity. Comparison of mass spectrometry spectra of sulfated glycans in 294-1B1-positive versus -negative OVCAR-3 cells indicated that the 294-1B1 epitope is likely at least 2, and possibly 3 or more, tandem GlcNAc-6-O-sulfated LacNAc units. Then, using the 294-1B1 antibody, we performed quantitative immunohistochemical analysis of 40 specimens from patients with ovarian cancer, consisting of 10 each of serous, endometrioid, clear cell, and mucinous carcinomas, and found that among them low-sulfated keratan sulfate was widely expressed in all but mucinous ovarian carcinoma.","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9831767","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Deglycosylated RBD produced in Pichia pastoris as a low-cost sera COVID-19 diagnosis tool and a vaccine candidate. 毕赤酵母生产的去糖基化RBD是一种低成本的血清新冠肺炎诊断工具和候选疫苗。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwad089

Tommy Idrovo-Hidalgo, María F Pignataro, Luis M Bredeston, Fernanda Elias, María G Herrera, María F Pavan, Sabrina Foscaldi, Mayra Suireszcz, Natalia B Fernández, Diana E Wetzler, Carlos H Paván, Patricio O Craig, Ernesto A Roman, Lucas A M Ruberto, Diego G Noseda, Lorena I Ibañez, Cecilia Czibener, Juan E Ugalde, Alejandro D Nadra, Javier Santos, Cecilia D'Alessio

{"title":"Deglycosylated RBD produced in Pichia pastoris as a low-cost sera COVID-19 diagnosis tool and a vaccine candidate.","authors":"Tommy Idrovo-Hidalgo, María F Pignataro, Luis M Bredeston, Fernanda Elias, María G Herrera, María F Pavan, Sabrina Foscaldi, Mayra Suireszcz, Natalia B Fernández, Diana E Wetzler, Carlos H Paván, Patricio O Craig, Ernesto A Roman, Lucas A M Ruberto, Diego G Noseda, Lorena I Ibañez, Cecilia Czibener, Juan E Ugalde, Alejandro D Nadra, Javier Santos, Cecilia D'Alessio","doi":"10.1093/glycob/cwad089","DOIUrl":"10.1093/glycob/cwad089","url":null,"abstract":"During the COVID-19 outbreak, numerous tools including protein-based vaccines have been developed. The methylotrophic yeast Pichia pastoris (synonymous to Komagataella phaffii) is an eukaryotic cost-effective and scalable system for recombinant protein production, with the advantages of an efficient secretion system and the protein folding assistance of the secretory pathway of eukaryotic cells. In a previous work, we compared the expression of SARS-CoV-2 Spike Receptor Binding Domain in P. pastoris with that in human cells. Although the size and glycosylation pattern was different between them, their protein structural and conformational features were indistinguishable. Nevertheless, since high mannose glycan extensions in proteins expressed by yeast may be the cause of a nonspecific immune recognition, we deglycosylated RBD in native conditions. This resulted in a highly pure, homogenous, properly folded and monomeric stable protein. This was confirmed by circular dichroism and tryptophan fluorescence spectra and by SEC-HPLC, which were similar to those of RBD proteins produced in yeast or human cells. Deglycosylated RBD was obtained at high yields in a single step, and it was efficient in distinguishing between SARS-CoV-2-negative and positive sera from patients. Moreover, when the deglycosylated variant was used as an immunogen, it elicited a humoral immune response ten times greater than the glycosylated form, producing antibodies with enhanced neutralizing power and eliciting a more robust cellular response. The proposed approach may be used to produce at a low cost, many antigens that require glycosylation to fold and express, but do not require glycans for recognition purposes.","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72014128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Correction to: ``MeCP2 regulated glycogenes contribute to proliferation and apoptosis of gastric cancer cells''. 更正：MeCP2调控糖原有助于胃癌细胞的增殖和凋亡

IF 4.3 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwad101

引用次数: 0

Lewis a-b- histo-blood group antigen phenotype is predictive of severe COVID-19 in the black South African population group. Lewis a-b-组织-血液组抗原表型可预测南非黑人群体中严重的新冠肺炎。

IF 3.4 3区生物学

Glycobiology Pub Date : 2024-03-19 DOI: 10.1093/glycob/cwad090

Cliff A Magwira, Ndivho P Nndwamato, Gloria Selabe, Mapaseka L Seheri

{"title":"Lewis a-b- histo-blood group antigen phenotype is predictive of severe COVID-19 in the black South African population group.","authors":"Cliff A Magwira, Ndivho P Nndwamato, Gloria Selabe, Mapaseka L Seheri","doi":"10.1093/glycob/cwad090","DOIUrl":"10.1093/glycob/cwad090","url":null,"abstract":"Several risk factors have been associated with SARS-CoV-2 infections and severity of COVID-19 disease it causes. This study investigated whether variations in histo-blood group antigen (HBGA) expression can predispose individuals to SARS-CoV-2 infections and severity of the disease. Nasopharyngeal swabs, randomly selected from SARS-CoV-2 positive and SARS-CoV-2 negative individuals, were tested for Lewis and H-type 1 HBGA phenotypes by ELISA using monoclonal antibodies specific to Lewis a, Lewis b and H type 1 antigens. The most common Lewis HBGA phenotype among all study participants was Lewis a-b+ (46%), followed by Lewis a-b- (24%), Lewis a+b- and Lewis a+b+ (15% each), while 55% of the study participants were H-type 1. Although SARS-CoV-2 negative individuals had a lower likelihood of having a Lewis a-b- phenotype compared to their SARS-CoV-2 positives counterparts (OR: 0.53, 95% C.I: 0.255-1.113), it did not reach statistical significance (P = 0.055). The frequency of Lewis a+b+, Lewis a+B-, Lewis a-b+, H type 1 positive and H type 1 negative were consistent between SARS-CoV-2 positive and SARS-CoV-2 negative individuals. When stratified according to severity of the disease, individuals with Lewis a+b- phenotype had a higher likelihood of developing mild COVID-19 symptoms (OR: 3.27, 95% CI; 0.9604-11.1), but was not statistically significant (P = 0.055), while Lewis a-b- phenotype was predictive of severe COVID-19 symptoms (OR: 4.3, 95% CI: 1.274-14.81), P = 0.016. In conclusion, individuals with Lewis a-b- phenotype were less likely to be infected by SARS-CoV-2, but when infected, they were at risk of severe COVID-19.","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72209069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0