Document Detail

Detection and characterization of a sialoglycosylated bacterial ABC-type phosphate transporter protein from patients with visceral leishmaniasis.
MedLine Citation:
PMID:  19184417     Owner:  NLM     Status:  MEDLINE    
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-O-AcSGPs) 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 alpha2-->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.
Angana Ghoshal; Sumi Mukhopadhyay; Rodion Demine; Michael Forgber; Saulius Jarmalavicius; Bibhuti Saha; Shyam Sundar; Peter Walden; Chhabinath Mandal; Chitra Mandal
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-01-29
Journal Detail:
Title:  Glycoconjugate journal     Volume:  26     ISSN:  1573-4986     ISO Abbreviation:  Glycoconj. J.     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-07-23     Completed Date:  2009-10-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8603310     Medline TA:  Glycoconj J     Country:  United States    
Other Details:
Languages:  eng     Pagination:  675-89     Citation Subset:  IM    
Department of Infectious Disease and Immunology, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Kolkata, 700032, India.
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MeSH Terms
Bacterial Proteins / chemistry*,  isolation & purification*
Chromatography, High Pressure Liquid
Chromatography, Thin Layer
Flow Cytometry
Leishmaniasis, Visceral / blood*
Membrane Transport Proteins / chemistry*,  isolation & purification*
Middle Aged
Molecular Weight
Sialoglycoproteins / chemistry*,  isolation & purification*
Spectrometry, Mass, Electrospray Ionization
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Young Adult
Reg. No./Substance:
0/Bacterial Proteins; 0/Membrane Transport Proteins; 0/Sialoglycoproteins

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