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Detection and characterization of a sialoglycosylated bacterial ABC-type phosphate transporter protein from patients with visceral leishmaniasis

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Title Detection and characterization of a sialoglycosylated bacterial
ABC-type phosphate transporter protein from patients
with visceral leishmaniasis
 
Creator Ghoshal, Angana
Mukhopadhyay, Sumi
Demine, Rodion
Forgber, Michael
Jarmalavicius, Saulius
Saha, Bibhuti
Sundar, Shyam
Walden, Peter
Mandal, Chhabinath
Mandal, Chitra
 
Subject Infectious Diseases and Immunology
Structural Biology & Bioinformatics
 
Description We report the discovery and characterization of a
glycosylated bacterial ABC-type phosphate transporter isolated
from the peripheral blood mononuclear cell (PBMC)
fraction of patients with visceral leishmaniasis (VL). Three
disease-associated 9-O-acetylated sialoglycoproteins (9-OAcSGPs)
of 19, 56 and 65 kDa, respectively, had been
identified and their purity, apparent mass and pI established
by SDS-PAGE and isoelectric focusing. Western blot
analyses showed that the 9-O-acetylated sialic acid is linked
via α2→6 linkage to a subterminal N-acetylgalactosamine.
For the 56 kDa protein, N- as well as O-glycosylations
were demonstrated by specific glycosidase treatment and
found to account for more than 9 kDa of the protein mass.
The presence of sialic acids was further confirmed through
thin layer chromatography, fluorimetric HPLC and electrospray ionization-mass spectrometry. The protein was
identified by mass spectrometry and de novo sequencing
of five tryptic fragments as a periplasmic ABC-type
phosphate transporter of Pseudomonas aeruginosa. The
amino acid sequences of the assigned peptides had 83–
100% identity with the NCBI entry for a Pseudomonas
transporter protein. Based on the recently reported X-ray
structure of a human phosphate-binding protein, we
predicted a 3D structural model for the 56 kDa protein
using homology and threading methods. The most probable
N- and O-glycosylation sites were identified by combinations
of sequence motif-searching bioinformatics tools, solvent
accessibility calculations, structural environment analyses and
mass spectrometric data. This is the first reported glycosylation
as well as sialylation of the periplasmic component of an
ABC-type phosphate transporter protein and of one of few
identified bacterial glycoproteins
 
Publisher Kluwer
 
Date 2009
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/173/1/GLYCOCONJUGATE_JOURNAL%2C__26(_6)%2C675%2D689%2C2009[41].pdf
Ghoshal, Angana and Mukhopadhyay, Sumi and Demine, Rodion and Forgber, Michael and Jarmalavicius, Saulius and Saha, Bibhuti and Sundar, Shyam and Walden, Peter and Mandal, Chhabinath and Mandal, Chitra (2009) Detection and characterization of a sialoglycosylated bacterial ABC-type phosphate transporter protein from patients with visceral leishmaniasis. Glycoconjugate Journal, 26 (6). pp. 675-689.
 
Relation http://dx.doi.org/10.1007/s10719-008-9223-8
http://www.eprints.iicb.res.in/173/