Sialylation of lactosyl lipids in membrane microdomains by T.cruzi trans-sialidase

Gavin Noble; Faye Craven; MD Segarra-Maset; Juana Reyes Martinez; Robert Sardzik; Sabine L Flitsch; SJ Webb

doi:10.1039/C4OB01852D

Sialylation of lactosyl lipids in membrane microdomains by T.cruzi trans-sialidase

Gavin Noble, Faye Craven, MD Segarra-Maset, Juana Reyes Martinez, Robert Sardzik, Sabine L Flitsch, SJ Webb

Research output: Contribution to journal › Article › peer-review

Abstract

A synthetic perfluoroalkyl-tagged lactosyl glycolipid has been shown to form lipid microdomains in fluid phospholipid bilayers. When embedded in the membranes of phospholipid vesicles, this glycolipid was trans-sialylated by soluble T. cruzi trans-sialidase (TcTS) to give a perfluoroalkyl-tagged glycolipid that displayed the ganglioside GM3 epitope, with up to 35% trans-sialylation from fetuin after 18 h. Following sialylation, vesicles bearing this Neu5Ac(α2-3)Gal(β1-4)Glc sequence in their “glycocalyx” were recognised and agglomerated by the lectin M. amurensis leukoagglutinin. Monitoring TcTS-mediated trans-sialylation by HPLC over the first 6 h revealed that enzymatic transformation of bilayer-embedded substrate was much slower than that of a soluble lactosyl substrate. Furthermore, clustering of the lactose-capped glycolipid into “acceptor” microdomains did not increase the rate of sialic acid transfer from fetuin by soluble TcTS, instead producing slight inhibition.

Original language	English
Pages (from-to)	9272-9278
Number of pages	6
Journal	Organic & biomolecular chemistry
Volume	12
Issue number	45
DOIs	https://doi.org/10.1039/C4OB01852D
Publication status	Published - 7 Dec 2014

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title = "Sialylation of lactosyl lipids in membrane microdomains by T.cruzi trans-sialidase",

abstract = "A synthetic perfluoroalkyl-tagged lactosyl glycolipid has been shown to form lipid microdomains in fluid phospholipid bilayers. When embedded in the membranes of phospholipid vesicles, this glycolipid was trans-sialylated by soluble T. cruzi trans-sialidase (TcTS) to give a perfluoroalkyl-tagged glycolipid that displayed the ganglioside GM3 epitope, with up to 35% trans-sialylation from fetuin after 18 h. Following sialylation, vesicles bearing this Neu5Ac(α2-3)Gal(β1-4)Glc sequence in their “glycocalyx” were recognised and agglomerated by the lectin M. amurensis leukoagglutinin. Monitoring TcTS-mediated trans-sialylation by HPLC over the first 6 h revealed that enzymatic transformation of bilayer-embedded substrate was much slower than that of a soluble lactosyl substrate. Furthermore, clustering of the lactose-capped glycolipid into “acceptor” microdomains did not increase the rate of sialic acid transfer from fetuin by soluble TcTS, instead producing slight inhibition.",

author = "Gavin Noble and Faye Craven and MD Segarra-Maset and {Reyes Martinez}, Juana and Robert Sardzik and Flitsch, {Sabine L} and SJ Webb",

note = "We thank Prof. A. C. C. Frasch, Universidad Nacional General San Mart{\'i}n, Argentina, for kindly providing the TcTS clone. J.E.R.M. thanks CONACyT-M{\'e}xico for funding (scholarship reference 214433/309655). F.C. and G.T.N. thank the BBSRC for providing studentship funding.",

year = "2014",

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doi = "10.1039/C4OB01852D",

language = "English",

volume = "12",

pages = "9272--9278",

journal = "Organic & biomolecular chemistry",

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T1 - Sialylation of lactosyl lipids in membrane microdomains by T.cruzi trans-sialidase

AU - Noble, Gavin

AU - Craven, Faye

AU - Segarra-Maset, MD

AU - Reyes Martinez, Juana

AU - Sardzik, Robert

AU - Flitsch, Sabine L

AU - Webb, SJ

N1 - We thank Prof. A. C. C. Frasch, Universidad Nacional General San Martín, Argentina, for kindly providing the TcTS clone. J.E.R.M. thanks CONACyT-México for funding (scholarship reference 214433/309655). F.C. and G.T.N. thank the BBSRC for providing studentship funding.

PY - 2014/12/7

Y1 - 2014/12/7

N2 - A synthetic perfluoroalkyl-tagged lactosyl glycolipid has been shown to form lipid microdomains in fluid phospholipid bilayers. When embedded in the membranes of phospholipid vesicles, this glycolipid was trans-sialylated by soluble T. cruzi trans-sialidase (TcTS) to give a perfluoroalkyl-tagged glycolipid that displayed the ganglioside GM3 epitope, with up to 35% trans-sialylation from fetuin after 18 h. Following sialylation, vesicles bearing this Neu5Ac(α2-3)Gal(β1-4)Glc sequence in their “glycocalyx” were recognised and agglomerated by the lectin M. amurensis leukoagglutinin. Monitoring TcTS-mediated trans-sialylation by HPLC over the first 6 h revealed that enzymatic transformation of bilayer-embedded substrate was much slower than that of a soluble lactosyl substrate. Furthermore, clustering of the lactose-capped glycolipid into “acceptor” microdomains did not increase the rate of sialic acid transfer from fetuin by soluble TcTS, instead producing slight inhibition.

AB - A synthetic perfluoroalkyl-tagged lactosyl glycolipid has been shown to form lipid microdomains in fluid phospholipid bilayers. When embedded in the membranes of phospholipid vesicles, this glycolipid was trans-sialylated by soluble T. cruzi trans-sialidase (TcTS) to give a perfluoroalkyl-tagged glycolipid that displayed the ganglioside GM3 epitope, with up to 35% trans-sialylation from fetuin after 18 h. Following sialylation, vesicles bearing this Neu5Ac(α2-3)Gal(β1-4)Glc sequence in their “glycocalyx” were recognised and agglomerated by the lectin M. amurensis leukoagglutinin. Monitoring TcTS-mediated trans-sialylation by HPLC over the first 6 h revealed that enzymatic transformation of bilayer-embedded substrate was much slower than that of a soluble lactosyl substrate. Furthermore, clustering of the lactose-capped glycolipid into “acceptor” microdomains did not increase the rate of sialic acid transfer from fetuin by soluble TcTS, instead producing slight inhibition.

U2 - 10.1039/C4OB01852D

DO - 10.1039/C4OB01852D

M3 - Article

SN - 1477-0520

VL - 12

SP - 9272

EP - 9278

JO - Organic & biomolecular chemistry

JF - Organic & biomolecular chemistry

IS - 45

ER -

Sialylation of lactosyl lipids in membrane microdomains by T.cruzi trans-sialidase

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