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

1999 ◽  
Vol 337 (3) ◽  
pp. 551-558 ◽  
Author(s):  
Abhijit GHOSH ◽  
Keya BANDYOPADHYAY ◽  
Labanyamoy KOLE ◽  
Pijush K. DAS
Keyword(s):  
Leishmania Donovani ◽  
Biochemical Characterization ◽  
Binding Protein ◽  
Outer Membrane Proteins ◽  
Deae Cellulose ◽  
Integral Membrane Protein ◽  
Mononuclear Phagocytes ◽  
Laminin Receptor ◽  
Laminin Binding Protein ◽  
Laminin Binding

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 specific receptor for laminin, a major ECM protein, with a Kd in the nanomolar range. Here we describe the identification, purification and biochemical characterization of the Leishmania laminin receptor. When the outer membrane proteins of L. donovani were blotted on to nitrocellulose paper and probed with laminin, a prominent laminin-binding protein of 67 kDa was identified. The purified 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 monospecific antibody. The same protein was obtained when parasite membrane extracts were adsorbed to antibody affinity matrix and eluted with glycine. The affinity-purified protein bound to laminin in a detergent-solubilized form as well as after integration into artificial 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 Leishmaniato host tissues.

scholarly journals Identification of a Treponema pallidum Laminin-Binding Protein

2003 ◽  
Vol 71 (5) ◽  
pp. 2525-2533 ◽  
Author(s):  
Caroline E. Cameron
Keyword(s):  
Binding Protein ◽  
Recombinant Expression ◽  
Outer Membrane Proteins ◽  
Treponema Pallidum ◽  
Binding Potential ◽  
Type I ◽  
Dependent Manner ◽  
Sodium Metaperiodate ◽  
Laminin Binding Protein ◽  
Laminin Binding

ABSTRACT Host extracellular matrix (ECM) components represent ideal microbial adhesion targets that many pathogens use for colonization of tissues and initiation of infection. This study investigated the interaction of the spirochete Treponema pallidum with the ECM component laminin. To identify candidate laminin-binding adhesins, the T. pallidum genome was analyzed to predict open reading frames that encode putative outer membrane proteins, as these proteins interact directly with host ECM components. Subsequent recombinant expression of these proteins and analysis of their laminin-binding potential identified one protein, Tp0751, that demonstrated specific attachment to laminin. Tp0751 attached to laminin in a dose-dependent, saturable manner but did not attach to the ECM component collagen type I or IV or to the negative control proteins fetuin or bovine serum albumin. Sodium metaperiodate treatment of laminin reduced the Tp0751-laminin interaction in a concentration-dependent manner, suggesting that oligosaccharides play a role in this interaction. In addition, Tp0751-specific antibodies were detected in serum samples collected from both experimental and natural syphilis infections, indicating that Tp0751 is expressed in vivo during the course of infection. Collectively, these experiments identified Tp0751 as a laminin-binding protein that is expressed during infection and may be involved in attachment of T. pallidum to host tissues.

Differential expression of the 67-kD laminin receptor and 31-kD human laminin-binding protein in human ovarian carcinomas

1994 ◽  
Vol 30 (8) ◽  
pp. 1096-1099 ◽  
Author(s):  
F.A. van den Brûle ◽  
A. Berchuck ◽  
R.C. Bast ◽  
Fu-Tong Liu ◽  
C. Gillet ◽  
...  
Keyword(s):  
Differential Expression ◽  
Binding Protein ◽  
Laminin Receptor ◽  
Ovarian Carcinomas ◽  
Laminin Binding Protein ◽  
Laminin Binding

scholarly journals Amino acid sequence and distribution of mRNA encoding a major skeletal muscle laminin binding protein: an extracellular matrix-associated protein with an unusual COOH-terminal polyaspartate domain.

1988 ◽  
Vol 107 (2) ◽  
pp. 699-705 ◽  
Author(s):  
D O Clegg ◽  
J C Helder ◽  
B C Hann ◽  
D E Hall ◽  
L F Reichardt
Keyword(s):  
Extracellular Matrix ◽  
Amino Acid ◽  
Membrane Protein ◽  
Amino Acid Sequence ◽  
Biochemical Characterization ◽  
Muscle Protein ◽  
Binding Protein ◽  
Single Gene ◽  
Laminin Binding Protein ◽  
Laminin Binding

Two cDNAs encoding an abundant chicken muscle extracellular matrix (ECM)-associated laminin-binding protein (LBP) have been isolated and sequenced. The predicted primary amino acid sequence includes a probable signal peptide and a site for N-linked glycosylation, but lacks a hydrophobic segment long enough to span the membrane. The COOH terminus consists of an unusual repeat of 33 consecutive aspartate residues. Comparison with other sequences indicates that this protein is different from previously described LBPs and ECM receptors. RNA blot analysis of LBP gene expression showed that LBP mRNA was abundant in skeletal and heart muscle, but barely detectable in other tissues. Blots of chicken genomic DNA suggest that a single gene encodes this LBP. The amino acid sequence and mRNA distribution are consistent with the biochemical characterization described by Hall and co-workers (Hall, D. E., K. A. Frazer, B. C. Hahn, and L. F. Reichardt. 1988. J. Cell Biol. 107:687-697). These analyses indicate that LBP is an abundant ECM-associated muscle protein with an unusually high negative charge that interacts with both membranes and laminin, and has properties of a peripheral, not integral membrane protein. Taken together, our studies show that muscle LBP is a secreted, peripheral membrane protein with an unusual polyaspartate domain. Its laminin and membrane binding properties suggest that it may help mediate muscle cell interactions with the extracellular matrix. We propose the name "aspartactin" for this LBP.

Expression and characterization of the ”laminin binding protein" in hydra

1997 ◽  
Vol 287 (3) ◽  
pp. 507-512 ◽  
Author(s):  
H. C. Schaller ◽  
E. Keppel ◽  
U. Fenger
Keyword(s):  
Binding Protein ◽  
Laminin Binding Protein ◽  
Laminin Binding
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