scholarly journals Identification of glycans on plasma-derived ADAMTS13

Blood ◽  
2016 ◽  
Vol 128 (21) ◽  
pp. e51-e58 ◽  
Author(s):  
Fabian C. Verbij ◽  
Eva Stokhuijzen ◽  
Paul H. P. Kaijen ◽  
Floris van Alphen ◽  
Alexander B. Meijer ◽  
...  

Key Points ADAMTS13 contains complex type N-linked glycans, which contain terminal mannose, sialic acids, and fucose residues. TSP1 repeats are modified by O-fucosylation and C-mannosylation; O-fucosylation was also observed in the disintegrin domain.

Biomolecules ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 136
Author(s):  
David Stenitzer ◽  
Réka Mócsai ◽  
Harald Zechmeister ◽  
Ralf Reski ◽  
Eva L. Decker ◽  
...  

In the animal kingdom, a stunning variety of N-glycan structures have emerged with phylogenetic specificities of various kinds. In the plant kingdom, however, N-glycosylation appears to be strictly conservative and uniform. From mosses to all kinds of gymno- and angiosperms, land plants mainly express structures with the common pentasaccharide core substituted with xylose, core α1,3-fucose, maybe terminal GlcNAc residues and Lewis A determinants. In contrast, green algae biosynthesise unique and unusual N-glycan structures with uncommon monosaccharides, a plethora of different structures and various kinds of O-methylation. Mosses, a group of plants that are separated by at least 400 million years of evolution from vascular plants, have hitherto been seen as harbouring an N-glycosylation machinery identical to that of vascular plants. To challenge this view, we analysed the N-glycomes of several moss species using MALDI-TOF/TOF, PGC-MS/MS and GC-MS. While all species contained the plant-typical heptasaccharide with no, one or two terminal GlcNAc residues (MMXF, MGnXF and GnGnXF, respectively), many species exhibited MS signals with 14.02 Da increments as characteristic for O-methylation. Throughout all analysed moss N-glycans, the level of methylation differed strongly even within the same family. In some species, methylated glycans dominated, while others had no methylation at all. GC-MS revealed the main glycan from Funaria hygrometrica to contain 2,6-O-methylated terminal mannose. Some mosses additionally presented very large, likewise methylated complex-type N-glycans. This first finding of the methylation of N-glycans in land plants mirrors the presumable phylogenetic relation of mosses to green algae, where the O-methylation of mannose and many other monosaccharides is a common trait.


Author(s):  
Mari A. Piirainen ◽  
Heidi Salminen ◽  
Alexander D. Frey

Abstract N-glycosylation is an important posttranslational modification affecting the properties and quality of therapeutic proteins. Glycoengineering in yeast aims to produce proteins carrying human-compatible glycosylation, enabling the production of therapeutic proteins in yeasts. In this work, we demonstrate further development and characterization of a glycoengineering strategy in a Saccharomyces cerevisiae Δalg3 Δalg11 strain where a truncated Man3GlcNAc2 glycan precursor is formed due to a disrupted lipid-linked oligosaccharide synthesis pathway. We produced galactosylated complex-type and hybrid-like N-glycans by expressing a human galactosyltransferase fusion protein both with and without a UDP-glucose 4-epimerase domain from Schizosaccharomyces pombe. Our results showed that the presence of the UDP-glucose 4-epimerase domain was beneficial for the production of digalactosylated complex-type glycans also when extracellular galactose was supplied, suggesting that the positive impact of the UDP-glucose 4-epimerase domain on the galactosylation process can be linked to other processes than its catalytic activity. Moreover, optimization of the expression of human GlcNAc transferases I and II and supplementation of glucosamine in the growth medium increased the formation of galactosylated complex-type glycans. Additionally, we provide further characterization of the interfering mannosylation taking place in the glycoengineered yeast strain. Key points • Glycoengineered Saccharomyces cerevisiae can form galactosylated N-glycans. • Genetic constructs impact the activities of the expressed glycosyltransferases. • Growth medium supplementation increases formation of target N-glycan structure.


1988 ◽  
Vol 263 (18) ◽  
pp. 8879-8889 ◽  
Author(s):  
L Roux ◽  
S Holojda ◽  
G Sundblad ◽  
H H Freeze ◽  
A Varki

2021 ◽  
Author(s):  
David Stenitzer ◽  
Réka Mócsai ◽  
Harald Zechmeister ◽  
Ralf Reski ◽  
Eva L. Decker ◽  
...  

In the animal kingdom, a stunning variety of N-glycan structures has emerged with phylogenetic specificities of various kinds. In the plant kingdom, however, N-glycosylation appears as strictly conservative and uniform. From mosses to all kinds of gymno- and angiosperms, land plants mainly express structures with the common pentasaccharide core substituted with xylose, core α1,3-fucose, maybe terminal GlcNAc residues and Lewis A determinants. In contrast, green algae biosynthesize unique and unusual N-glycan structures with uncommon monosaccharides, a plethora of different structures and various kinds of O-methylation. Mosses, a group of plants that are separated by at least 400 million years of evolution from vascular plants, were hitherto seen as harbouring an N-glycosylation machinery identical to that of vascular plants. To challenge this view, we have analysed the N-glycomes of several moss species using MALDI-TOF/TOF, PGC-MS/MS and GC-MS. While all species contained the plant-typical heptasaccharide with no, one or two terminal GlcNAc residues (MMXF, MGnXF and GnGnXF, respectively), many species exhibited MS signals with 14.02 Da increments as characteristic for O-methylation. Throughout all analysed moss N-glycans the level of methylation differed strongly even in the same family. In some species, methylated glycans dominated, while others had no methylation at all. GC-MS revealed the main glycan from Funaria hygrometrica to contain 2,6-O-methylated terminal mannose. Some mosses additionally presented very large, likewise methylated complex-type N-glycans. This first finding of methylation of N-glycans in land plants mirrors the presumable phylogenetic relation of mosses to green algae, where O-methylation of mannose and many other monosaccharides is a common trait.


Blood ◽  
2016 ◽  
Vol 127 (22) ◽  
pp. 2701-2710 ◽  
Author(s):  
Satu Hyvärinen ◽  
Seppo Meri ◽  
T. Sakari Jokiranta

Key Points Sialic acids are critical for factor H–mediated complement regulation on endothelial cells, erythrocytes, and platelets. Impaired ability of factor H mutants to simultaneously bind sialic acid and C3b on cells explains their association with aHUS.


Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3617 ◽  
Author(s):  
Tamas Pongracz ◽  
Manfred Wuhrer ◽  
Noortje de Haan

The human glycome is characterized by a high degree of sialylation, affecting, amongst others, cell–cell interactions and protein half-life. An established method for the linkage isomer-specific characterization of N-glycan sialylation is based on the linkage-specific derivatization of sialylated glycoconjugates, inducing ethyl esterification of α2,6-linked sialic acids and lactonization of α2,3-linked sialic acids. While the carboxylic acid activator and nucleophile used in this reaction received extensive investigation, the role of the catalyst was never thoroughly explored. A frequently used catalyst for the linkage-specific esterification of sialic acids is 1-hydroxybenzotriazole (HOBt). Here, a systematic evaluation was performed of five HOBt alternatives in combination with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) in ethanol for the linkage-specific derivatization of sialic acids. Derivatized glycans were analyzed by MALDI-TOF-MS and the catalyst performance was evaluated based on the completeness of the reactions and the linkage-specificity obtained. The use of both 6-Cl-HOBt and 6-CF3-HOBt resulted in high linkage-specificity and minimal byproduct formation, similar to the benchmark method using HOBt. Performing the reaction with these catalysts at neutral or acidic pH showed comparable efficiencies on both sialyllactose and complex-type N-glycans. The reported investigations resulted in an expansion of the reaction space for linkage-specific sialic acid derivatization.


1981 ◽  
Vol 195 (3) ◽  
pp. 691-699 ◽  
Author(s):  
T Tsuji ◽  
K Yamamoto ◽  
T Irimura ◽  
T Osawa

The unit A-type glycopeptides were purified from porcine thyroglobulin by Pronase digestion followed by chromatography on a DEAE-Sephadex A-25 column. These glycopeptides were separated into five fractions (UA-I, -II, -IV and -V) by Dowex 50W (X2) column chromatography. Fractions UA-I, -II, -III, -IV and -V were found to have the compositions (Man)9(GlcNAc)2-Asn, (Man)8(GlcNAc)2-Asn, (Man)7(GlcNAc)2-Asn, (Man)6(GlcNAc)2-Asn and (Man)5(GlcNAc)2-Asn respectively. The structures of these five fractions were investigated by the combination of exo- and endo-glycosidase digestions, methylation analysis. Smith periodate degradation and acetolysis. The results showed that fraction UA-V had the simplest structure: see formula in text. The larger glycopeptides (fractions UA-I, -II, -III and -IV) contained additional mannose residues alpha (1 leads to 2)-linked to the terminal mannose residues in the above core structure. These unit A-type glycopeptides appear to be biosynthetic intermediates that are to be processed to form complex-type glycopeptides (unit B-type sugar chains).


Author(s):  
Laura S. DeThorne ◽  
Kelly Searsmith

Purpose The purpose of this article is to address some common concerns associated with the neurodiversity paradigm and to offer related implications for service provision to school-age autistic students. In particular, we highlight the need to (a) view first-person autistic perspectives as an integral component of evidence-based practice, (b) use the individualized education plan as a means to actively address environmental contributions to communicative competence, and (c) center intervention around respect for autistic sociality and self-expression. We support these points with cross-disciplinary scholarship and writings from autistic individuals. Conclusions We recognize that school-based speech-language pathologists are bound by institutional constraints, such as eligibility determination and Individualized Education Program processes that are not inherently consistent with the neurodiversity paradigm. Consequently, we offer examples for implementing the neurodiversity paradigm while working within these existing structures. In sum, this article addresses key points of tension related to the neurodiversity paradigm in a way that we hope will directly translate into improved service provision for autistic students. Supplemental Material https://doi.org/10.23641/asha.13345727


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