Molecular Cloning and Expression of the Mouse N-Acetylneuraminic Acid 9-Phosphate Synthase Which Does Not Have Deaminoneuraminic Acid (KDN) 9-Phosphate Synthase Activity

Nakata, Daisuke; Close, Brett E.; Colley, Karen J.; Matsuda, Tsukasa; Kitajima, Ken

doi:10.1006/bbrc.2000.2983

Cited by 31 publications

(28 citation statements)

References 33 publications

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“…The primers, based on the mouse adipo Q nucleotide sequence (GenBank TM accession number U49915) used were 5Ј-GTTGGATGGCAGGCATCC-3Ј (nucleotides 122-140) and 5Ј-CCTG-GAGCCAGACTTGGTC-3Ј (nucleotides 634 -652). 3Ј-and 5Ј-rapid amplification of cDNA ends were carried out as described previously (22). Nucleotide sequences of the cloned cDNAs were determined by the dideoxy chain termination method (23) on a DNA sequencer model 373 (Applied Biosystems).…”

Section: Methodsmentioning

confidence: 99%

Identification and Adipocyte Differentiation-dependent Expression of the Unique Disialic Acid Residue in an Adipose Tissue-specific Glycoprotein, Adipo Q

Sato¹,

Yasukawa²,

Honda³

et al. 2001

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Identification and Adipocyte Differentiation-dependent Expression of the Unique Disialic Acid Residue in an Adipose Tissue-specific Glycoprotein, Adipo Q

Sato¹,

Yasukawa²,

Honda³

et al. 2001

Journal of Biological Chemistry

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“…cDNA Cloning of WGA16 -Preparation of total RNA and oligo(dT)-derived cDNA from pig male reproductive organs (testis, epididymis, seminal vesicles, prostate, and bulbourethral glands) and rapid amplification of cDNA ends (RACE) were performed as described previously (30). Based on a partial amino acid sequence that was identified as homologous to ZG16 homolog B from S. scrofa, the cDNA fragment for WGA16 was amplified by reverse transcription polymerase chain reaction (RT-PCR) from prostate and bulbourethral glands using the following primers: ZG16-F-P (5Ј-CAGATGT-TCGGGAACGGAAAAGGCTCC-3Ј) and ZG16-R-P (5Ј-TATAATGTGCTCGCCGGGNTGCAGGAT-3Ј).…”

Section: Sds-page/lectin Blotting and Westernmentioning

confidence: 99%

Discovery, Primary, and Crystal Structures and Capacitation-related Properties of a Prostate-derived Heparin-binding Protein WGA16 from Boar Sperm

Garenáux

Kanagawa

Tsuchiyama

et al. 2015

Journal of Biological Chemistry

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Background: Mechanisms of glycoprotein redistribution during sperm capacitation remain unclear. Results: Prostate-specific ZG16-like lectin WGA16 was discovered in sperm lipid rafts. Its primary and crystal structure and GalT-and heparin-binding properties were characterized. Conclusion: Attachment to sperm via surface GalT and capacitation-induced detachment involve unique N-glycans and the heparin-binding domain. Significance: This is the first demonstration of a glycan-mediated mechanism for transient existence of seminal plasma glycoprotein on the sperm surface.

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“…This plasmid had a unique EcoRI site within the v-cath open reading frame, which was used to insert an in-frame E. coli LacZ fragment encoding a functional β-galactosidase protein and produce a new baculovirus transfer plasmid, pAcVcath-LacRI. In parallel, we excised a KpnI fragment encoding murine SAS (Nakata et al, 2000) and CMP-SAS (Munster et al, 1998) under the transcriptional control of dual AcMNPV ie1 promoters and the AcMNPV hr5 enhancer element from p64KDIE1TV1/SAS/CMP-SAS and then blunted the ends and subcloned it into the unique EcoRI site (blunted) of p64KTV1ΔK/NΔS/P to produce another baculovirus transfer plasmid, pAcSWT-7. Both of these new transfer plasmids were isolated from large-scale bacterial cultures by standard alkaline lysis and CsCl-ethidium bromide density gradient ultracentrifugation methods, as described previously (Sambrook et al, 1989), and their genetic structures were confirmed by restriction mapping.…”

Section: Construction Of Baculovirus Transfer Plasmidsmentioning

confidence: 99%

“…AcSWT-7B encodes two murine enzymes, SAS (Nakata et al, 2000) and CMP-SAS (Munster et al, 1998), which can convert N-acetylmannosamine through a sialic acid intermediate to the nucleotide sugar, CMP-sialic acid (Fig. 1).…”

Section: Isolation Of a Recombinant Baculovirus Encoding Murine Cmp-smentioning

confidence: 99%

Isolation and analysis of a baculovirus vector that supports recombinant glycoprotein sialylation by SfSWT‐1 cells cultured in serum‐free medium

Hill¹,

Aumiller²,

Shi³

et al. 2006

Biotech & Bioengineering

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The inability to sialylate recombinant glycoproteins is a critical limitation of the baculovirusinsect cell expression system. This limitation is due, at least in part, to the absence of detectable sialyltransferase activities and CMP-sialic acids in the insect cell lines routinely used as hosts in this system. SfSWT-1 is a transgenic insect cell line encoding five mammalian glycosyltransferases, including sialyltransferases, which can contribute to sialylation of recombinant glycoproteins expressed by baculovirus vectors. However, sialylation of recombinant glycoproteins requires culturing SfSWT-1 cells in the presence of fetal bovine serum or another exogenous source of sialic acid. To eliminate this requirement and extend the utility of SfSWT-1 cells, we have isolated a new baculovirus vector, AcSWT-7B, designed to express two mammalian enzymes that can convert N-acetylmannosamine to CMP-sialic acid during the early phase of infection. AcSWT-7B was also designed to express a model recombinant glycoprotein during the very late phase of infection. Characterization of this new baculovirus vector showed that it induced high levels of intracellular CMP-sialic acid and sialylation of the recombinant Nglycoprotein upon infection of SfSWT-1 cells cultured in serum-free medium supplemented with N-acetylmannosamine. In addition, co-infection of SfSWT-1 cells with AcSWT-7B plus a conventional baculovirus vector encoding human tissue plasminogen activator resulted in sialylation of this recombinant N-glycoprotein under the same culture conditions. These results demonstrate that AcSWT-7B can be used in two different ways to support recombinant Nglycoprotein sialylation by SfSWT-1 cells in serum-free medium. Thus, AcSWT-7B can be used to extend the utility of this previously described transgenic insect cell line for recombinant sialoglycoprotein production.

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Molecular Cloning and Expression of the Mouse N-Acetylneuraminic Acid 9-Phosphate Synthase Which Does Not Have Deaminoneuraminic Acid (KDN) 9-Phosphate Synthase Activity

Cited by 31 publications

References 33 publications

Identification and Adipocyte Differentiation-dependent Expression of the Unique Disialic Acid Residue in an Adipose Tissue-specific Glycoprotein, Adipo Q

Identification and Adipocyte Differentiation-dependent Expression of the Unique Disialic Acid Residue in an Adipose Tissue-specific Glycoprotein, Adipo Q

Discovery, Primary, and Crystal Structures and Capacitation-related Properties of a Prostate-derived Heparin-binding Protein WGA16 from Boar Sperm

Isolation and analysis of a baculovirus vector that supports recombinant glycoprotein sialylation by SfSWT‐1 cells cultured in serum‐free medium

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