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Isolation of a laminin-binding protein from the protozoan parasite Leishmania donovani that may mediate cell adhesion

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Title Isolation of a laminin-binding protein from the protozoan parasite
Leishmania donovani that may mediate cell adhesion
 
Creator Ghosh, Abhijit
Bandyopadhyay, Keya
Kole, Labanyamoy
Das, Pijush K
 
Subject Infectious Diseases and Immunology
 
Description Extracellular matrix (ECM)-binding proteins on the surface of
Leishmania are thought to play a crucial role in the onset of
leishmaniasis, as these parasites invade mononuclear phagocytes
in various organs after migrating through the ECM. In a previous
report, we presented several lines of evidence suggesting that
Leishmania has a speci®c receptor for laminin, a major ECM
protein, with a Kd in the nanomolar range. Here we describe the
identi®cation, puri®cation and biochemical characterization of
the Leishmania laminin receptor. When the outer membrane
proteins of L. dono�ani were blotted on to nitrocellulose paper
and probed with laminin, a prominent laminin-binding protein
of 67 kDa was identi®ed. The puri®ed protein was isolated by a
three-step process involving DEAE±cellulose, Con A (concanavalin
A)±Sepharose and laminin±Sepharose affinity chromatography
and was used to raise a monospeci®c antibody. The
same protein was obtained when parasite membrane extracts were adsorbed to antibody affinity matrix and eluted with glycine.
The affinity-puri®ed protein bound to laminin in a detergentsolubilized
form as well as after integration into arti®cial bilayers,
and was subsequently characterized as an integral membrane
protein. Metaperiodate oxidation and metabolic inhibition of
glycosylation studies indicate the binding protein to be glycoprotein
in nature and that N-linked oligosaccharides play a
part in the interaction of laminin with the binding protein.
Surface-labelled parasites attached to microtitre wells coated
with laminin and the 67 kDa protein blocked the adhesion to
laminin substrate. We propose that the 67 kDa protein is an
adhesin involved in the attachment of Leishmania to host tissues.
 
Publisher Portland Press
 
Date 1999
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/654/1/BIOCHEMICAL_JOURNAL_337%2C_551%2D558;1999[77].pdf
Ghosh, Abhijit and Bandyopadhyay, Keya and Kole, Labanyamoy and Das, Pijush K (1999) Isolation of a laminin-binding protein from the protozoan parasite Leishmania donovani that may mediate cell adhesion. Biochemical Journal, 337. pp. 551-558. ISSN 0264-6021
 
Relation http://dx.doi.org/10.1042/0264-6021:3370551
http://www.eprints.iicb.res.in/654/