Colorimetric assay of sialic acid by a methyl-3-benzothiazolinone-2-hydrazone reactant

Massamiri, Y.; Beljean, M.; Durand, Geneviève; Pays, M.; Agneray, J

doi:10.1016/0003-2697(78)90548-1

Cited by 14 publications

(5 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The sugar derivatives were detected by the UV absorbance of the benzoyl group at 230 nm, and the retention times were within the range from 20 to 50 min. In addition, TV-acetylneuraminic acid was determined by a colorimetric method (Massamiri et al, 1978). No de-O-acetylation of samples was performed prior to analysis, for which reason only TV-acetylneuraminic acid, but not 9-0and 8-O-acetylated forms, was determined.…”

Section: Methodsmentioning

confidence: 99%

“…No de-O-acetylation of samples was performed prior to analysis, for which reason only TV-acetylneuraminic acid, but not 9-0and 8-O-acetylated forms, was determined. Human -1-acid glycoprotein (Sigma) was used as standard assuming a value of 9.48% (w/w) of sialic acid (Massamiri et al, 1978). For the calculation of moles of sialic acid per mole of factor VIIa protein, molecular weights of 309 and 45 514 were used for TV-acetylneuraminic acid and nonglycosylated factor VIIa, respectively.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells

Thim

Bjoern²,

Christensen³

et al. 1988

Biochemistry

290

186

View full text Add to dashboard Cite

Blood coagulation factor VII is a vitamin K dependent glycoprotein which in its activated form, factor VIIa, participates in the coagulation process by activating factor X and/or factor IX in the presence of Ca2+ and tissue factor. Three types of potential posttranslational modifications exist in the human factor VIIa molecule, namely, 10 gamma-carboxylated, N-terminally located glutamic acid residues, 1 beta-hydroxylated aspartic acid residue, and 2 N-glycosylated asparagine residues. In the present study, the amino acid sequence and posttranslational modifications of recombinant factor VIIa as purified from the culture medium of a transfected baby hamster kidney cell line have been compared to human plasma factor VIIa. By use of HPLC, amino acid analysis, peptide mapping, and automated Edman degradations, the protein backbone of recombinant factor VIIa was found to be identical with human factor VIIa. Neither recombinant factor VIIa nor human plasma factor VIIa was found to contain beta-hydroxyaspartic acid. In human plasma factor VIIa, the 10 N-terminally located glutamic acid residues were found to be fully gamma-carboxylated whereas 9 full and 1 partial gamma-carboxylated residues were found in the corresponding positions of the recombinant factor VIIa molecule. Asparagine residues 145 and 322 were found to be fully N-glycosylated in human plasma factor VIIa. In the recombinant factor VIIa, asparagine residue 322 was fully glycosylated whereas asparagine residue 145 was only partially (approximately 66%) glycosylated. Besides minor differences in the sialic acid and fucose contents, the overall carbohydrate compositions were nearly identical in recombinant factor VIIa and human plasma factor VIIa.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells

Thim

Bjoern²,

Christensen³

et al. 1988

Biochemistry

290

186

View full text Add to dashboard Cite

show abstract

“…In this study, we employed the MBTHmethod (Massamiri et al -1978) to quantitatively determine total, free and bound sialic acids in the luminal fluid of different regions in the rat and hamster epididymis. T h s method has been used previously to determine bound sialic acid in the rat epididymal fluid (Toowicharanont -1982) as well as on the rat epididymal sperm surface (Toowicharanont and Chulavatnatol -1983).…”

Section: Discussionmentioning

confidence: 99%

“…In this study, we employed the MBTHmethod (Massamiri et al -1978) to quantitatively determine total, free and bound sialic 25.6 f 0.9…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Free and Bound Sialic Acid in Rat and Hamster Epididymal Fluid/Freie und gebundene Sialinsäure in der Nebenhodenflüssigkeit der Ratte und des Hamsters

Verawatnapakul

Pholpramool

2009

Andrologia

View full text Add to dashboard Cite

Total, free and bound sialic'acid concentrations were determined in sperm-free luminal fluid removed by micropuncture from different regions of the rat and hamster epididymides. In the rat, total sialic acid concentrations (mean f s.e.m.) in the proximal caput, the mid corpus and the proximal cauda were, respectively, 25.7 f 1.3, 23.9 f 1.7 and 28.8 f 1.9 mM compared to 4.4 f 0.1 mM in blood plasma. In the hamster, total sialic acid concentrations in the distal corpus, the proximal cauda and the distal cauda were, respectively, 32.9 f 3.8, 26.4 f 1.4 and 26.6 f 3.2 mM compared to 4.7 f 0.5 mM in blood plasma. Free sialic acid accounted for approximately 7(t80%0 of total sialic acid present in the epididymal plasma of the rat. Similarly, 82% of sialic acid in the rat blood plasma was in free form. The levels of free and bound sialic acid were not changed in different regions of the rat epididymis. Unilateral Ligation of the rat efferent ducts had no effect on total, free or bound sialic acid concentrations in all regions except in the proximal cauda in which a transient increase (P < 0.01) in free sialic acid was observed on Day-3 after EDL. Zusammenfassung: Im spermatozoenfreien Sekret des Ratten-und Hamsternebenhodens wurden Gesamt -, freie und gebundene -Sialinsaure bestimmt, nachdem das Sekret durch Mikropunktion gewonnen war. Bei der Ratte ergaben sich unterschiedliche Konzentrationen fir Gesamtsialinsaure (GS) in den verschiedenen Nebenhodenabschnitten (Kopf: 25.7 f 1.3 mM, Mittelteil: 23.9 f 1.7 mM, Schwanz: 28.8 f 1.9 mM) gegenuber einer Konzentration von 4.4 f 0.1 mM im Blutplasma. Beim Hamster ergab sich folgendes: Distaler Corpus: 32.9 f 3.8 mM; proximale Cauda: 26.4 f 1.4 mM, distale Cauda: 26.6 f 3.2 mM gegenuber einer Konzentration von 4.7 f 0.5 m M i m Blutplasma.Freie Sialinsaure wurde fir ungefsihr 70430% der Gesamtsialinsaure berechnet, die im Nebenhodenplasma der Ratte vorhanden war. In 5hnlicher Weise lag bei der Ratte 82% der Sialinsaure i m Blutplasma in freier Form vor. In den verschiedenen Abschnitten des Rattennebenhodens ergaben sich keine Veranderungen der Werte fir freie und gebundene Sialinsaure. Eine einseitige Ligatur der Ductus efferentes zeigte bei der Ratte keinerlei EinfluB auf die Werte f~ Gesamt-, sowie freie oder gebundene Sialinsaure mit Ausnahme der proximalen Cauda, bei der ein voriibergehender Anstieg (p <0.01) an freier Sialinsaure am dritten postoperativen Tage beobachtet werden konnte. andrologia 20, No. 5 (1988)

show abstract

Expression of PNA‐binding sites on specific glycoproteins by human melanoma cells is associated with a high metastatic potential

Zebda

Bailly

Brown

et al. 1994

J of Cellular Biochemistry

View full text Add to dashboard Cite

Lectin-binding patterns of seven human melanoma clones and variants selected from the same parental cell line and differing in their spontaneous metastatic potential in an animal model were compared by flow cytometry and Scatchard analysis. Human melanoma clones and variants with high and low metastatic potential could be distinguished by their peanut agglutinin (PNA)-binding patterns, but not by their wheat germ agglutinin (WGA)-, Ulex europaeus agglutinin I (UEA I)-, and soybean agglutinin (SBA)-binding patterns. Low metastatic clones and variants proved to be made up of single poorly peanut agglutinin-binding cell population (2.20-3.52 x 10(6) sites/cell, Ka = 2.48-2.75 x 10(6) M-1). By contrast, highly metastatic variants were found to be constituted by two cellular subpopulations, exhibiting respectively a moderate 2.62-3.72 x 10(6) sites/cell) and a high peanut agglutinin staining (17.68-18.76 x 10(6) sites/cell). One highly metastatic clone was found to be homogeneously constituted by a single population of cells strongly binding this lectin (18.86 x 10(6) sites/cell) with an association constant of 4.06 +/- 10(6) M-1. Using an EPICS V cytometer, these two subpopulations were sorted from a highly metastatic variant and tested for their metastatic abilities: cells with high PNA binding generated a higher frequency of metastases than did moderately PNA-binding cells. Following treatment with Vibrio cholerae neuraminidase, all cells from all variants and clones were brightly labeled by PNA, collecting in a single peak with similar fluorescence intensities. Electrophoresis of total cellular proteins and subsequent detection with labeled PNA om Western blots show two major PNA-reactive glycoproteins with apparent molecular weights of 140 and 110 kDa (MAGP1 and MAGP2), expressed only in highly metastatic cells, but which can be strongly labeled by PNA in slightly metastatic cells following a treatment with neuraminidase. These results provide evidence that the expression of terminal galactose (beta 1-3)N-acetyl galactosamine structure, positioned on MAGP1 and MAGP2 glycoproteins, is associated with the metastatic potential of human melanoma cells.

show abstract

Colorimetric assay of sialic acid by a methyl-3-benzothiazolinone-2-hydrazone reactant

Cited by 14 publications

References 6 publications

Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells

Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells

Free and Bound Sialic Acid in Rat and Hamster Epididymal Fluid/Freie und gebundene Sialinsäure in der Nebenhodenflüssigkeit der Ratte und des Hamsters

Expression of PNA‐binding sites on specific glycoproteins by human melanoma cells is associated with a high metastatic potential

Contact Info

Product

Resources

About