Purification of the Porcine rubulavirus attachment protein by liquid isoelectric focusing

Santos‐López, Gerardo; Flores, Esmeralda; Baños, Rocı́o; Herrera‐Camacho, Irma; Reyes‐Leyva, Julio

doi:10.1016/j.pep.2003.12.009

Cited by 10 publications

(7 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Neuraminidase activity was determined at 37 °C in sodium acetate buffer (0.1 M , pH 4.5) using bovine fetuin as substrate. Sialic acid released by influenza viruses was quantified colorimetrically by a modification of the thiobarbituric acid method 59. Neu N PhAc and Neu N Ac displayed similar response to this colorimetric assay (see Supporting Information S2.3).…”

Section: Resultsmentioning

confidence: 98%

Chemoenzymatic Syntheses of Sialylated Oligosaccharides Containing C5‐Modified Neuraminic Acids for Dual Inhibition of Hemagglutinins and Neuraminidases

Birikaki

Pradeau

Armand

et al. 2015

Chemistry A European J

Self Cite

View full text Add to dashboard Cite

A fast chemoenzymatic synthesis of sialylated oligosaccharides containing C5-modified neuraminic acids is reported. Analogues of GM3 and GM2 ganglioside saccharidic portions where the acetyl group of NeuNAc has been replaced by a phenylacetyl (PhAc) or a propanoyl (Prop) moiety have been efficiently prepared with metabolically engineered E. coli bacteria. GM3 analogues were either obtained by chemoselective modification of biosynthetic N-acetyl-sialyllactoside (GM3 NAc) or by direct bacterial synthesis using C5-modified neuraminic acid precursors. The latter strategy proved to be very versatile as it led to an efficient synthesis of GM2 analogues. These glycomimetics were assessed against hemagglutinins and sialidases. In particular, the GM3 NPhAc displayed a binding affinity for Maackia amurensis agglutinin (MAA) similar to that of GM3 NAc, while being resistant to hydrolysis by Vibrio cholerae (VC) neuraminidase. A preliminary study with influenza viruses also confirmed a selective inhibition of N1 neuraminidase by GM3 NPhAc, suggesting potential developments for the detection of flu viruses and for fighting them.

show abstract

Section: Resultsmentioning

confidence: 98%

Chemoenzymatic Syntheses of Sialylated Oligosaccharides Containing C5‐Modified Neuraminic Acids for Dual Inhibition of Hemagglutinins and Neuraminidases

Birikaki

Pradeau

Armand

et al. 2015

Chemistry A European J

Self Cite

View full text Add to dashboard Cite

show abstract

“…Indeed, their predicted pI differed significantly: 7.79 for HN-K 335 versus 6.99 for HN-E 335 . However, we found that purified HN-K 335 and HN-E 335 proteins presented pIs of 4.30 and 4.65, respectively (data not shown), similar to porcine rubulavirus HN protein (pI 4.4) and other neuraminidases [35]. This difference between theoretical and experimental values could be because predicted pI is only based on The first column presents the function associated with each set of residues.…”

Section: Electrostatic Potential Analysis Of Hn-k 335 and Hn-e 335 Mumentioning

confidence: 91%

“…Data from references: [11,27,28,[30][31][32][44][45][46][47]. amino acid sequences, although it can be modified in nature by glycosylation, as has been demonstrated for other viral glycoproteins [35,36]. Since, both HN-K 335 and HN-E 335 proteins preserved the seven potential N-glycosylation sites (Figure 2), it is probable that as for other MuV strains, at least five are actually occupied by carbohydrate sidechains that reduce the pI values [37,38].…”

Section: Electrostatic Potential Analysis Of Hn-k 335 and Hn-e 335 Mumentioning

confidence: 99%

Structure-function analysis of two variants of mumps virus hemagglutinin-neuraminidase protein

Santos‐López¹,

Scior²,

Borraz-Argüello³

et al. 2009

Braz J Infect Dis

Self Cite

View full text Add to dashboard Cite

A point mutation from guanine (G) to adenine (A) at nucleotide position 1081 in the hemagglutinin-neuraminidase (HN) gene has been associated with neurovirulence of Urabe AM9 mumps virus vaccine. This mutation corresponds to a glutamic acid (E) to lysine (K) change at position 335 in the HN glycoprotein. We have experimentally demonstrated that two variants of Urabe AM9 strain (HN-A 1081 and HN-G 1081 ) differ in neurotropism, sialic acidbinding affinity and neuraminidase activity. In the present study, we performed a structure-function analysis of that amino acid substitution; the structures of HN protein of both Urabe AM9 strain variants were predicted. Based on our analysis, the E/K mutation changes the protein surface properties and to a lesser extent their conformations, which in turn reflects in activity changes. Our modeling results suggest that this E/K interchange does not affect the structure of the sialic acid binding motif; however, the electrostatic surface differs drastically due to an exposed short alpha helix. Consequently, this mutation may affect the accessibility of HN to substrates and membrane receptors of the host cells. Our findings appear to explain the observed differences in neurotropism of these vaccine strains.

show abstract

“…Se demostró que la HN del RVP es una proteína altamente termoestable, hecho que ya se había explorado para la cepa LPM (Reyes-Leyva et al 1999, Santos-López et al 2004b. Es importante apuntar que PAC1 mostró un nivel más alto de resistencia a la temperatura, lo cual también sugiere que su HN posee caracterís-ticas diferentes a las de LPM y PAC3, que le permiten mantener un mínimo de su estructura funcional después del tratamiento térmico.…”

Section: Discussionunclassified

“…Los virus purificados o sobrenadantes virales se incubaron en presencia de sialil-lactosa (Sigma) o fetuína sérica bovina (Sigma) como sustratos a 37 ºC en una solución amortiguadora de acetato de sodio 0.1 M (Merck, Darmstad, Alemania) a pH 3.5 durante 30 min. El ácido neuramínico liberado se determinó mediante un microensayo modificado del método del ácido tiobarbitúrico, usando una curva de ácido neuramínico puro, de acuerdo con el protocolo publicado previamente (Santos-López et al 2004b).…”

Section: Ensayos De Hemaglutinaciónunclassified

Caracterización biológica de tres aislamientos naturales del Rubulavirus porcino (México)

Borraz-Argüello¹,

Santos‐López²,

Vallejo-Ruíz³

et al. 2007

RBT

View full text Add to dashboard Cite

Biological characterization of three natural isolates of the porcine rubulavirus (Mexico).Porcine rubulavirus (PoRV) produces a neurological and reproductive syndrome in pigs called the blue-eye disease, known only from Mexico. Several isolates were grouped by the main symptoms presented during outbreaks: a) neurotropic in piglets, b) broadly neurotropic in piglets and gonadotropic in adults, and c) gonadotropic in adults. We studied some biological properties of three strains, which fall in one of each virus group: La Piedad Michoacán (LPM) and Producción Animal Cerdos 1 (PAC1) and 3 (PAC3), respectively. The analyzed viral properties are mainly related with the trans-membrane hemagglutinin-neuraminidase (HN) and fusion (F) proteins, such as cytopathic effect, hemolysis, hemagglutinating (HA) and neuraminidase (NA) activities. In the infection assays PAC1 strain presented the highest fusogenicity level; however, the most cytolytic strain was PAC3. In addition, HA and NA activities and viral genome of PAC3 strain was detected in supernatants during cell infection earlier than in the other two strains, which shows that PAC3 virions release from the host cell earlier than LPM and PAC1. Experimental determination in purified viruses shows that PAC3 presented a higher HA and NA activities; however, PAC1 shows other interesting properties, such as a high thermostability of HN and differences about substrate profile respect to LPM and PAC3. Our data suggest that NA activity is associated with the virulence of RVP. Rev. Biol. Trop. 56 (2): 487-499. Epub 2008 June 30.

show abstract

Purification of the Porcine rubulavirus attachment protein by liquid isoelectric focusing

Cited by 10 publications

References 30 publications

Chemoenzymatic Syntheses of Sialylated Oligosaccharides Containing C5‐Modified Neuraminic Acids for Dual Inhibition of Hemagglutinins and Neuraminidases

Chemoenzymatic Syntheses of Sialylated Oligosaccharides Containing C5‐Modified Neuraminic Acids for Dual Inhibition of Hemagglutinins and Neuraminidases

Structure-function analysis of two variants of mumps virus hemagglutinin-neuraminidase protein

Caracterización biológica de tres aislamientos naturales del Rubulavirus porcino (México)

Contact Info

Product

Resources

About