scholarly journals A highly conserved surface loop in the C-terminal domain of ovotransferrin (residues 570-584) is remote from the receptor-binding site

1990 ◽  
Vol 266 (2) ◽  
pp. 393-398 ◽  
Author(s):  
A B Mason ◽  
S A Brown ◽  
W R Church
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
Polyclonal Antibodies ◽  
Receptor Binding Site ◽  
Cellular Receptors ◽  
Phase Synthesis ◽  
Surface Loop ◽  
Terminal Domain

A peptide corresponding to a surface loop in the C-terminal domain of chicken ovotransferrin (residues 570-584) was made by solid-phase synthesis and used to immunize rabbits. A 15-amino acid-residue disulphide-linked loop occurs in both domains of all five transferrins for which the sequence is available and lies on the opposite side of the iron-binding site from the interdomain cleft. Polyclonal antibodies to the peptide were specific for non-reduced holo-ovotransferrin and the C-terminal domain, as shown by e.l.i.s.a. and immunoblotting. The antibody did not inhibit binding of ovotransferrin to receptors on chick-embryo reticulocytes but was able to bind ovotransferrin bound to the cellular receptors at 0 degree C. The loop composed of residues 570-584 appears to be remote from the transferrin receptor-binding site.

scholarly journals Inhibition of the specific binding of human lactotransferrin to human peripheral-blood phytohaemagglutinin-stimulated lymphocytes by fluorescein labelling and location of the binding site

1991 ◽  
Vol 276 (3) ◽  
pp. 733-738 ◽  
Author(s):  
D Legrand ◽  
J Mazurier ◽  
P Maes ◽  
E Rochard ◽  
J Montreuil ◽  
...  
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Specific Binding ◽  
Peripheral Blood Lymphocytes ◽  
Receptor Binding Site ◽  
Terminal Domain ◽  
Total Fluorescence ◽  
Domain I

Labelling of human lactotransferrin with fluorescein 5′-isothiocyanate (FITC) in an equimolar ratio inhibits the binding of the protein to phytohaemagglutinin-activated human peripheral-blood lymphocytes. Therefore it can be assumed that FITC reacts at, or near, the receptor-binding site. Three FITC-labelled peptides have been purified from a tryptic digest of the FITC-labelled lactotransferrin. The determination of their amino acid sequence and their localization on the primary structure of the protein permitted the identification of two FITC-accessible areas in the N-terminal lobe and one in the C-terminal lobe. In fact, only 10% of the total FITC was conjugated to one lysine residue (Lys579) of the C-terminal lobe, whereas most (80%) of the FITC was conjugated to three close lysine residues [Lys263 (65% of total fluorescence), Lys280 and Lys282 (15% of total fluorescence)] located in beta-turn structures, of the N-terminal domain I of human lactotransferrin. The results obtained show that the receptor-binding site should be located in the vicinity of the FITC-accessible Lys263, Lys280 and Lys282, and corroborate our preliminary results reporting the involvement of the N-terminal domain I in the binding of human lactotransferrin to mitogen-stimulated lymphocytes [Rochard, Legrand, Mazurier, Montreuil & Spik (1989) FEBS Lett. 255, 201-204]. In any case, FITC labelling is not suitable for studying the binding of lactotransferrin to activated lymphocytes and its use may lead to erroneous interpretations of cell binding experiments.

Divergent solid-phase synthesis and candidacidal activity of MUC7 D1, a 51-residue histidine-rich N-terminal domain of human salivary mucin MUC7

2000 ◽  
Vol 56 (5) ◽  
pp. 275-282 ◽  
Author(s):  
J. Satyanarayana ◽  
H. Situ ◽  
S. Narasimhamurthy ◽  
N. Bhayani ◽  
L.A. Bobek ◽  
...  
Keyword(s):  
Solid Phase Synthesis ◽  
Solid Phase ◽  
Phase Synthesis ◽  
Terminal Domain ◽  
Salivary Mucin

scholarly journals Crystal Structure of a Chimeric Receptor Binding Protein Constructed from Two Lactococcal Phages

2009 ◽  
Vol 191 (10) ◽  
pp. 3220-3225 ◽  
Author(s):  
Marina Siponen ◽  
Silvia Spinelli ◽  
Stéphanie Blangy ◽  
Sylvain Moineau ◽  
Christian Cambillau ◽  
...  
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Domain Adaptation ◽  
Host Recognition ◽  
Diverse Population ◽  
Positive Bacterium ◽  
Receptor Binding Site ◽  
Terminal Domain ◽  
Linker Domain ◽  
Slight Displacement

ABSTRACT Lactococcus lactis, a gram-positive bacterium widely used by the dairy industry to manufacture cheeses, is subject to infection by a diverse population of virulent phages. We have previously determined the structures of three receptor binding proteins (RBPs) from lactococcal phages TP901-1, p2, and bIL170, each of them having a distinct host range. Virulent phages p2 and bIL170 are classified within the 936 group, while the temperate phage TP901-1 is a member of the genetically distinct P335 polythetic group. These RBPs comprise three domains: the N-terminal domain, binding to the virion particle; a β-helical linker domain; and the C-terminal domain, bearing the receptor binding site used for host recognition. Here, we have designed, expressed, and determined the structure of an RBP chimera in which the N-terminal and linker RBP domains of phage TP901-1 (P335) are fused to the C-terminal RBP domain of phage p2 (936). This chimera exhibits a stable structure that closely resembles the parental structures, while a slight displacement of the linker made RBP domain adaptation efficient. The receptor binding site is structurally indistinguishable from that of native p2 RBP and binds glycerol with excellent affinity.

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