Exploration of Conformational Spaces of High-Mannose-Type Oligosaccharides by an NMR-Validated Simulation

SummaryModification of the carbohydrate structures of recombinant tissue-type plasminogen activator (rt-PA) can increase or decrease its rate of clearance in rabbits. When rt-PA was treated with sodium periodate to oxidize carbohydrate residues, the rate of clearance was decreased from 9.6 ± 1.9 ml min−1 kg−1 to 3.5 ± 0.6 ml min−1 kg−1 (mean ± SD, n = 5). A similar change in the clearance of rt-PA was introduced by the use of endo-β-N-acetyl- glucosaminidase H (Endo-H), which selectively removes high mannose asparagine-linked oligosaccharides; the clearance of Endo-H-treated rt-PA was 5.0 ± 0.5 ml min−1 kg−1. A mutant of rt-PA was produced with an amino acid substitution at position 117 (Asn replaced with Gin) to remove a potential glycosylation site that normally contains a high mannose structure. The clearance of this material was also decreased, similar to the periodate and Endo-H-treated rt-PA. Conversely, when rt-PA was produced in the CHO 15B cell line, which can produce only high mannose oligosaccharide structures on glycoproteins, the clearance was increased by a factor of 1.8. These results demonstrate that the removal of rt-PA from the blood depends significantly upon the nature of its oligosaccharide structures.

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One extra oligosaccharide chain of the high-mannose class in the E2 protein of a Sindbis virus isolate.

Journal of Virology ◽

10.1128/jvi.38.1.8-14.1981 ◽

1981 ◽

Vol 38 (1) ◽

pp. 8-14 ◽

Cited By ~ 11

Author(s):

R Cancedda ◽

S Bonatti ◽

A Leone

Keyword(s):

Sindbis Virus ◽

Virus Isolate ◽

E2 Protein ◽

Oligosaccharide Chain ◽

High Mannose

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Structural studies of the major high mannose oligosaccharide units from Chinese hamster ovary cell glycoproteins.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)37814-6 ◽

1979 ◽

Vol 254 (5) ◽

pp. 1600-1605 ◽

Cited By ~ 26

Author(s):

E. Li ◽

S. Kornfeld

Keyword(s):

Chinese Hamster Ovary Cell ◽

Chinese Hamster Ovary ◽

Chinese Hamster ◽

Ovary Cell ◽

Structural Studies ◽

High Mannose

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Control of glycoprotein synthesis. The purification by preparative high voltage paper electrophoresis in borate of glycopeptides containing high mannose and complex oligosaccharide chains linked to asparagine.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)85578-3 ◽

1980 ◽

Vol 255 (10) ◽

pp. 4876-4884

Author(s):

S. Narasimhan ◽

N. Harpaz ◽

G. Longmore ◽

J.P. Carver ◽

A.A. Grey ◽

...

Keyword(s):

High Voltage ◽

Paper Electrophoresis ◽

Glycoprotein Synthesis ◽

High Mannose

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BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms

Biomolecules ◽

10.3390/biom11020180 ◽

2021 ◽

Vol 11 (2) ◽

pp. 180

Author(s):

Zorana Lopandić ◽

Luka Dragačević ◽

Dragan Popović ◽

Uros Andjelković ◽

Rajna Minić ◽

...

Keyword(s):

Fluorescent Protein ◽

Lectin Binding ◽

Three Dimensional ◽

Data Bank ◽

Salmonella Typhi ◽

Excitation Wavelength ◽

Protein Data Bank Entry ◽

High Mannose

Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms.

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