Molecular Cloning of Siaα2,3Galβ1,4GlcNAc α2,8-Sialyltransferase from Mouse Brain

Yoshida, Yukiko; Kojima, Naoya; Kurosawa, Nobuyuki; Hamamoto, Toshiro; Tsuji, S.

doi:10.1074/jbc.270.24.14628

Cited by 99 publications

(74 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sialic acid residues are usually not extended further with other monosaccharides, except with another sialic acid. Sialyltransferases of the mammalian ST8Sia family catalyze oligo-and polysialylation of glycoproteins through transfer of α2,8-sialic acids from CMP-sialic acid to the nonreducing ends of α2,6-or α2,3-Sia acceptors (20,21). If the degree of polymerization (DP) exceeds 8 (DP >8), such structures are regarded as polySia chains and can be as long as DP >400 (3).…”

Section: Human Polysialyltransferases Are Functional When Transientlymentioning

confidence: 99%

Engineering of complex protein sialylation in plants

Kallolimath

Castilho

Strasser

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Sialic acids (Sias) are abundant terminal modifications of protein-linked glycans. A unique feature of Sia, compared with other monosaccharides, is the formation of linear homo-polymers, with its most complex form polysialic acid (polySia). Sia and polySia mediate diverse biological functions and have great potential for therapeutic use. However, technological hurdles in producing defined protein sialylation due to the enormous structural diversity render their precise investigation a challenge. Here, we describe a plant-based expression platform that enables the controlled in vivo synthesis of sialylated structures with different interlinkages and degree of polymerization (DP). The approach relies on a combination of stably transformed plants with transient expression modules. By the introduction of multigene vectors carrying the human sialylation pathway into glycosylation-destructed mutants, transgenic plants that sialylate glycoproteins in α2,6- or α2,3-linkage were generated. Moreover, by the transient coexpression of human α2,8-polysialyltransferases, polySia structures with a DP >40 were synthesized in these plants. Importantly, plant-derived polySia are functionally active, as demonstrated by a cell-based cytotoxicity assay and inhibition of microglia activation. This pathway engineering approach enables experimental investigations of defined sialylation and facilitates a rational design of glycan structures with optimized biotechnological functions.

show abstract

Section: Human Polysialyltransferases Are Functional When Transientlymentioning

confidence: 99%

Engineering of complex protein sialylation in plants

Kallolimath

Castilho

Strasser

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Our new findings further suggest that biosynthesis of these di-Sia and oligo-Sia residues on glycoproteins may be catalyzed by ␣238-sialyltransferase(s) distinct from the known polysialyltransferases, STX and PST, which are at least partially responsible for polysialylation of N-CAM (40 -42). In this regard, ST8SiaIII, whose acceptor substrate has not yet been identified (43), may be a candidate enzyme for synthesis of these novel glycotopes.…”

Section: Discussionmentioning

confidence: 99%

Frequent Occurrence of Pre-existing α2→8-Linked Disialic and Oligosialic Acids with Chain Lengths Up to 7 Sia Residues in Mammalian Brain Glycoproteins

Sato

Fukuoka

Ohta

et al. 2000

Journal of Biological Chemistry

107

View full text Add to dashboard Cite

The pre-existence of ␣238-linked disialic acid (di-Sia) and oligosialic acid (oligo-Sia) structures with up to 7 Sia residues was shown to occur on a large number of brain glycoproteins, including neural cell adhesion molecules (N-CAMs), by two highly sensitive chemical methods (Sato, C., Inoue, S., Matsuda, T., and Kitajima, K. (1998) Anal. Biochem. 261, 191-197; Sato, C., Inoue, S., Matsuda, T., and Kitajima, K. (1999) Anal. Biochem. 266, 102-109). This unexpected finding was also confirmed using a newly developed antibody prepared using a copolymer of ␣238-linked N-acetylneuraminyl p-vinylbenzylamide and acrylamide as an immunogen and known antibodies whose immunospecificities were determined to be di-and oligo-Sia residues with defined chain lengths. The major significance of the new finding that di-and oligo-Sia chains exist on a large number of brain glycoproteins is 2-fold. First, it reveals a surprising diversity in the number and M r of proteins distinct from N-CAM that are covalently modified by these short sialyl glycotopes. Second, it suggests that synthesis of diand/or oligo-Sia units may be catalyzed by ␣238-sialyltransferase(s) that are distinct from the known polysialyltransferases, STX and PST, which are partially responsible for polysialylation of N-CAM.

show abstract

“…3). We previously showed that PSA was synthesized on fetuin by ST8Sia II and IV but not by ST8Sia III (11,13,21). The 14 C-sialylated polydisperse materials were synthesized in addition to 14 C-sialylated fetuin by ST8Sia II and IV, while such a polydisperse material was not synthesized from fetuin by ST8Sia III.…”

Section: Ncam Is Specifically Polysialylated In Vivo Onmentioning

confidence: 99%

Characterization of Mouse ST8Sia II (STX) as a Neural Cell Adhesion Molecule-specific Polysialic Acid Synthase

Kojima

Tachida

Yoshida

et al. 1996

Journal of Biological Chemistry

Self Cite

116

View full text Add to dashboard Cite

Polysialic acid (PSA) 1 is an unusual carbohydrate associated with the neural cell adhesion molecule (NCAM) and the ␣-subunit of the voltage-gated sodium channel in mammals and is more abundant in embryonic than adult brain (1-3). PSA is implicated in the reduction of NCAM adhesion through its large negative charge and thereby allows increased neurite outgrowth and cell motility (4 -6). It is also reported that the presence of PSA affects cell-cell interactions through other adhesion molecules (7). Although functions of PSA have been described, little is known about the mechanisms by which the synthesis and expression of PSA are regulated.PSA on NCAM has so far been considered to be synthesized through the actions of at least two distinct enzymes, i.e. an initiator ␣2,8-sialyltransferase, which adds a single ␣2,8-linked sialic acid to ␣2,3-linked sialic acid residues on N-glycans, and a polysialyltransferase, which adds multiple ␣2,8-linked sialic acid residues (8). Livingston and Paulson (9) cloned STX/ST8Sia II, a new member of the sialyltransferase gene family from rat. Recently, we showed that ST8Sia II/STX cloned from mouse can synthesize PSA on ␣2,3-sialylated Nglycans of glycoproteins without an initiator ␣2,8-sialyltransferase in vitro and direct in vivo synthesis of PSA in several cells (10,11). In addition, we cloned mouse ST8Sia IV by homology cloning using the sequence information on GD3 synthase (12) and STX (9) and showed that the mouse enzyme can synthesize PSA, like ST8Sia II/STX (13). Eckhardt et al. (14) also cloned a sialyltransferase, which is involved in the biosynthesis of PSA on NCAM from hamster, namely PST-1. Hamster PST-1 showed 99.2% identity to mouse ST8Sia IV, indicating that the hamster enzyme is a counterpart of mouse ST8Sia IV. The gene expression of mouse ST8Sia II was restricted in the brain and was dramatically up-and down-regulated during developmental stages of the brain like the expression of PSA, whereas mouse ST8Sia IV was strongly expressed in lung and heart rather than brain, and only little regulation was observed during brain development (10, 13). These observations suggest that ST8Sia II/STX rather than ST8Sia IV/PST-1 is involved in the biosynthesis of PSA associated with NCAM during mouse brain development. However, there is no evidence that the PSA on NCAM is specifically synthesized by ST8Sia II in vivo as well as in vitro, because PSA was synthesized on some glycoproteins such as fetuin in vitro (11), and the transfection of the human STX gene into NCAM-negative cells also caused the expression of PSA on the cell surface (15).The present study focuses on the biosynthesis of PSA by mouse ST8Sia II using a recombinant soluble NCAM fused with the Fc region of human IgG1 (NCAM-Fc) and comparison of the activities of mouse ST8Sia II and IV. ST8Sia II specifically and directly synthesized PSA on ␣2,3-linked sialic acid residues of N-glycans of specific isoforms of NCAM, without

show abstract

Molecular Cloning of Siaα2,3Galβ1,4GlcNAc α2,8-Sialyltransferase from Mouse Brain

Cited by 99 publications

References 27 publications

Engineering of complex protein sialylation in plants

Engineering of complex protein sialylation in plants

Frequent Occurrence of Pre-existing α2→8-Linked Disialic and Oligosialic Acids with Chain Lengths Up to 7 Sia Residues in Mammalian Brain Glycoproteins

Characterization of Mouse ST8Sia II (STX) as a Neural Cell Adhesion Molecule-specific Polysialic Acid Synthase

Contact Info

Product

Resources

About