Comparative proteomics and glycoproteomics of plasma proteins in Indian visceral leishmaniasisReport as inadecuate




Comparative proteomics and glycoproteomics of plasma proteins in Indian visceral leishmaniasis - Download this document for free, or read online. Document in PDF available to download.

Proteome Science

, 12:48

First Online: 22 September 2014Received: 17 May 2014Accepted: 08 September 2014

Abstract

BackgroundVisceral leishmaniasis VL is a deadly parasitic diseases caused by Leishmania donovani; it is a major health problem in many countries. A lack of proper understanding of the disease biology, poor diagnostic methods and increasing drug resistance are the main reasons for the growing burden of VL infection. Comparative plasma proteomics are a relatively useful technique that can be used to investigate disease-associated alterations that can help in understanding host responses against pathogens, and might be useful in disease management and diagnosis.

ResultIn this study, a comparative proteomics and glycoproteomics approach using 2DE and 2D-DIGE was employed between early diagnosed VL patients of all age groups and healthy endemic and non-endemic controls in order to aid the recognition of disease-associated alterations in host plasma. Comparative proteomics was performed by the depletion of seven highly abundant plasma proteins. Comparative glycoproteomics was performed by the depletion of albumin and IgG, followed by purification of plasma glycoproteins using a multi lectin affinity column. From these two approaches, 39 differentially expressed protein spots were identified and sequenced using MALDI-TOF-TOF mass spectrometry. This revealed ten distinct proteins that appeared in multiple spots, suggesting micro-heterogeneity. Among these proteins, alpha-1-antitrypsin, alpha-1-B glycoprotein and amyloid-A1 precursor were up-regulated, whereas vitamin-D binding protein, apolipoprotein-A-I and transthyretin were down-regulated in VL. Alterations in the levels of these proteins in VL-infected plasma were further confirmed by western blot and ELISA.

ConclusionsThese proteins may be involved in the survival of parasites, resisting neutrophil elastase, and in their multiplication in macrophages, potentially maintaining endogenous anti-inflammatory and immunosuppressive conditions. Consequently, the results of this study may help in understanding the host response against L.donovani, which could help in the discovery of new drugs and disease management. Finally, these alterations on protein levels might be beneficial in improving early diagnosis considering those as biomarkers in Indian VL.

KeywordsVisceral leishmaniasis Plasma glycoproteomics MARS column M-LAC column 2D-DIGE MALDI-TOF-TOF mass spectrometry AbbreviationsVLVisceral Leishmaniasis

2DETwo dimensional gel electrophoresis

2D-DIGETwo dimensional differential in-gel electrophoresis

IEFIsoelectric focusing

pIIsoelectric point

IPGImmobilized pH gradient

MALDI-TOF-TOFMatrix assisted laser desorption ionization-time of flight-time of flight

M-LACMulti lectin affinity chromatography, MARS, multiple affinity removal system

MWCOMolecular weight cut off

DTTDithiothreitol

TFATrifluoro-acetic acid

MSMass spectrometry

SDSSodium dodecyl sulphate

PAGEPolyacrylamide gel electrophoresis

CHAPS3-3-cholamidopropyl dimetylammonio-1-propanesulfonate

Electronic supplementary materialThe online version of this article doi:10.1186-s12953-014-0048-z contains supplementary material, which is available to authorized users.

Download fulltext PDF



Author: Arup Kumar Bag - Sutapa Saha - Shyam Sundar - Bibhuti Saha - Abhijit Chakrabarti - Chitra Mandal

Source: https://link.springer.com/







Related documents