scholarly journals Receptor binding of guinea pig and pig vasoactive intestinal peptides by rat lung

1988 ◽  
Vol 254 (2) ◽  
pp. 613-615 ◽  
Author(s):  
S Paul ◽  
D J Volle ◽  
J Currie
Keyword(s):  
Amino Acid ◽  
Guinea Pig ◽  
Vasoactive Intestinal Peptide ◽  
Receptor Binding ◽  
Amino Acid Residues ◽  
Rat Lung ◽  
Intestinal Peptides ◽  
Peptide Hydrolases ◽  
Vasoactive Intestinal Peptides

Guinea pig vasoactive intestinal peptide (gpVIP) differs from other mammalian VIPs in four of its 28 amino acid residues. In the present study, the gpVIP displaced 125I-labelled pig VIP (pVIP) binding by rat lung membranes with 7.7-fold lower potency than pVIP. Degradation of gpVIP by rat lung membranes, assessed by radioimmunoassay and h.p.l.c., was 1.9-fold greater than that of pVIP. This difference in degradation of the two peptides was not large enough to account for the lower receptor-binding potency of gpVIP. The amino acid residues that distinguish pVIP from gpVIP are likely to contribute to the interaction of VIP with receptors and peptide hydrolases in lung membranes.

scholarly journals Novel antibody binding determinants on the capsid surface of serotype O foot-and-mouth disease virus

2014 ◽  
Vol 95 (5) ◽  
pp. 1104-1116 ◽  
Author(s):  
Amin S. Asfor ◽  
Sasmita Upadhyaya ◽  
Nick J. Knowles ◽  
Donald P. King ◽  
David J. Paton ◽  
...  
Keyword(s):  
Amino Acid ◽  
Guinea Pig ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Amino Acid Residues ◽  
Serotype O ◽  
Antigenic Sites ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
The Impact

Five neutralizing antigenic sites have been described for serotype O foot-and-mouth disease viruses (FMDV) based on monoclonal antibody (mAb) escape mutant studies. However, a mutant virus selected to escape neutralization of mAb binding at all five sites was previously shown to confer complete cross-protection with the parental virus in guinea pig challenge studies, suggesting that amino acid residues outside the mAb binding sites contribute to antibody-mediated in vivo neutralization of FMDV. Comparison of the ability of bovine antisera to neutralize a panel of serotype O FMDV identified three novel putative sites at VP2-74, VP2-191 and VP3-85, where amino acid substitutions correlated with changes in sero-reactivity. The impact of these positions was tested using site-directed mutagenesis to effect substitutions at critical amino acid residues within an infectious copy of FMDV O1 Kaufbeuren (O1K). Recovered viruses containing additional mutations at VP2-74 and VP2-191 exhibited greater resistance to neutralization with both O1K guinea pig and O BFS bovine antisera than a virus that was engineered to include only mutations at the five known antigenic sites. The changes at VP2-74 and VP3-85 are adjacent to critical amino acids that define antigenic sites 2 and 4, respectively. However VP2-191 (17 Å away from VP2-72), located at the threefold axis and more distant from previously identified antigenic sites, exhibited the most profound effect. These findings extend our knowledge of the surface features of the FMDV capsid known to elicit neutralizing antibodies, and will improve our strategies for vaccine strain selection and rational vaccine design.

ReaxFF-based molecular dynamics simulation of the interaction between plasma ROS and the receptor-binding domain of the spike protein in the capsid protein of SARS-CoV-2

2021 ◽  
Author(s):  
Huichao Wang ◽  
Tong Zhao ◽  
Shuhui Yang ◽  
Liang Zou ◽  
Xiaolong Wang ◽  
...  
Keyword(s):  
Molecular Dynamics ◽  
Amino Acid ◽  
Capsid Protein ◽  
Receptor Binding ◽  
Hydrogen Abstraction ◽  
Binding Domain ◽  
Spike Protein ◽  
Receptor Binding Domain ◽  
Amino Acid Residues ◽  
Global Epidemic

Abstract Under the severe situation of the current global epidemic, researchers have been working hard to find a reliable way to suppress the infection of the virus and prevent the spread of the epidemic. Studies have shown that the recognition and binding of human angiotensin-converting enzyme 2 (ACE2) by the receptor-binding domain (BRD) of spike protein on the surface of SARS-CoV-2 is a crucial step for SARS-CoV-2 to invade human receptor cells, and blocking this process can inhibit the virus from invading human normal cells. Plasma treatment can disrupt the structure of the RBD and effectively block the binding process. However, the mechanism by which plasma blocks the recognition and binding between the two is not clear. In this study, reaction process between reactive oxygen species (ROS) in plasma and the molecular model of RBD was simulated using a reactive molecular dynamics method. The results showed that the destruction of RBD molecule by ROS was triggered by hydrogen abstraction reactions. O and OH abstracted H atoms from RBD, while the H atoms of H2O2 and HO2 were abstracted by RBD. The hydrogen abstraction resulted in the breakage of C-H, N-H, O-H and C=O bonds and the formation of C=C, C=N bonds. The addition reaction of OH increased the number of O-H bonds and caused the formation of C-O, N-O and O-H bonds. The dissociation of N-H bonds led to the destruction of the original structure of peptide bonds and amino acid residues, change the type of amino acid residues, and caused the conversion of N-C and N=C, C=O and C-O. The simulation partially elucidated the microscopic mechanism of the interaction between ROS in plasma and the capsid protein of SARS-CoV-2, providing theoretical support for the control of SARS-CoV-2 infection by plasma, a contribution to overcoming the global epidemic problem.

scholarly journals cDNA cloning and characterization of guinea-pig leukotriene B4 receptor

1999 ◽  
Vol 342 (1) ◽  
pp. 79-85 ◽  
Author(s):  
Kazuyuki MASUDA ◽  
Takehiko YOKOMIZO ◽  
Takashi IZUMI ◽  
Takao SHIMIZU
Keyword(s):  
Amino Acid ◽  
Guinea Pig ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Leukotriene B4 ◽  
Xenopus Laevis Oocytes ◽  
Amino Acid Residues ◽  
Ovary Cells ◽  
Hek 293 ◽  
Leukotriene B4 Receptor

The cDNA for leukotriene B4 (LTB4) receptor (BLT) was cloned from a guinea-pig leucocyte cDNA library. The cloned receptor cDNA encodes 348 amino acid residues and shares 73% identity with the amino acid sequence of human BLT. Northern blot analysis showed the highest expression of the receptor mRNA in leucocytes, followed by lung and spleen. The membrane fractions of HEK-293 and Cos-7 cells transfected with the cDNA showed specific LTB4-binding activities, with Kd values of 0.27 and 0.17 nM respectively. Xenopus laevis oocytes injected with the cRNA of guinea-pig BLT showed LTB4-induced Cl- currents, indicating that the cloned receptor is functional. LTB4 is metabolized to 20-hydroxy-LTB4 and then to 20-carboxy-LTB4, a transformation considered as a major inactivation pathway of the compound. Using the cloned receptor, we analysed the agonistic effects of LTB4 and these two metabolites. 20-Carboxy-LTB4 is a much weaker agonist, with a Kd value higher than that of LTB4 by three orders of magnitude, corresponding to a much weaker chemotactic activity. Although 20-hydroxy-LTB4 is as potent as LTB4 in inhibiting [3H]LTB4 binding and cAMP formation, it is less potent than LTB4 in the mobilization of intracellular Ca2+ and the chemotaxis of Chinese hamster ovary cells expressing the guinea-pig BLT. The present study demonstrated that although LTB4 and 20-hydroxy-LTB4 bind to the receptor with similar affinities, they do differ in activating intracellular signalling.

scholarly journals Receptor Binding Domain of Escherichia coli F18 Fimbrial Adhesin FedF Can Be both Efficiently Secreted and Surface Displayed in a Functional Form in Lactococcus lactis

2004 ◽  
Vol 70 (4) ◽  
pp. 2061-2071 ◽  
Author(s):  
Agneta Lindholm ◽  
Andreas Smeds ◽  
Airi Palva
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Receptor Binding ◽  
Intestinal Epithelial Cells ◽  
Surface Display ◽  
Cell Surface Display ◽  
Binding Domain ◽  
Intestinal Epithelial ◽  
Receptor Binding Domain ◽  
Amino Acid Residues

ABSTRACT Adherence of F18 fimbrial Escherichia coli to porcine intestinal epithelial cells is mediated by the adhesin (FedF) of F18 fimbriae. In a previous study, we demonstrated the specificity of the amino acid residues between 60 and 109 as the receptor binding domain of FedF. In this study, different expression, secretion, and anchoring systems for the receptor binding domain of the FedF adhesin in Lactococcus lactis were evaluated. Two partially overlapping receptor binding domains (42 and 62 amino acid residues) were expressed as fusions with L. lactis subsp. cremoris protein PrtP for evaluation of secretion efficiency. To evaluate the cell surface display of these FedF-PrtP fusions, they were further combined with different lengths of PrtP spacers fused with either the L. lactis AcmA anchor or the PrtP cell wall binding domain. An HtrA-defective L. lactis NZ9000 mutant was constructed to determine its effect on the level of secreted or anchored fusion proteins. Recombinant L. lactis clones secreting the receptor binding domain of F18 fimbriae as a fusion with the H domains of L. lactis protein PrtP were first constructed by using two different signal peptides. FedF-PrtP fusions, directed by the signal sequence of L. brevis SlpA, were throughout found to be secreted at significantly higher quantities than corresponding fusions with the signal peptide of L. lactis Usp45. In the surface display systems tested, the L. lactis AcmA anchor performed significantly better, particularly in the L. lactis NZ9000ΔhtrA strain, compared to the L. lactis PrtP anchor region. Of the cell surface display constructs with the AcmA anchor, only those with the longest PrtP spacer regions resulted in efficient binding of recombinant L. lactis cells to porcine intestinal epithelial cells. These results confirmed that it is possible to efficiently produce the receptor binding domain of the F18 adhesin in a functionally active form in L. lactis.

scholarly journals Binding of various ovotransferrin fragments to chick-embryo red cells

1989 ◽  
Vol 257 (1) ◽  
pp. 301-304 ◽  
Author(s):  
A Oratore ◽  
G D'Andrea ◽  
K Moreton ◽  
J Williams
Keyword(s):  
Amino Acid ◽  
Chick Embryo ◽  
Red Blood Cells ◽  
Receptor Binding ◽  
Blood Cells ◽  
Tight Binding ◽  
Red Cells ◽  
Transferrin Receptors ◽  
Amino Acid Residues

1. The ability of N- and C-terminal half-molecule fragments of hen ovotransferrin to interact with chick red blood cells (CERBC) has been studied under conditions that allow binding of the transferrin to transferrin receptors to take place, but not the delivery of iron to the cell. Two kinds of half-molecule fragments were used: (a) those which can associate with one another to give a dimer resembling native transferrin and (b) those which cannot associate in this way because they lack a few amino acid residues from their C-terminal ends. 2. Neither N nor C half-molecules alone can bind to the CERBC, but, when both are present, tight binding occurs. 3. Whether or not the half-molecules can associate with one another makes little difference to receptor binding. 4. Given that one of the half-molecules is iron-saturated, the presence or absence of iron in the contralateral half-molecule again makes little difference to receptor binding.

scholarly journals Primary structure of the cytosolic β-glucosidase of guinea pig liver

1996 ◽  
Vol 319 (3) ◽  
pp. 829-837 ◽  
Author(s):  
William S HAYS ◽  
Steven A. JENISON ◽  
Takashi YAMADA ◽  
Andrzej PASTUSZYN ◽  
Robert H. GLEW
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Guinea Pig ◽  
Mammalian Species ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
Pig Liver ◽  
Reading Frame ◽  
Degenerate Oligonucleotide ◽  
Guinea Pig Liver

The cytosolic β-glucosidase (EC 3.2.1.21) present in the livers of mammalian species is distinguished by its broad specificity for sugars and its preference for hydrophobic aglycones. We purified the cytosolic β-glucosidase from guinea pig liver and sequenced 142 amino acid residues contained within 12 trypsin digest fragments. Using degenerate oligonucleotide primers deduced from the peptide sequences, a 622 bp cytosolic β-glucosidase cDNA was amplified by reverse-transcriptase PCR, using total guinea pig liver RNA as template. The ‘rapid amplification of cDNA ends (RACE)’ method [Frohman (1993) Methods Enzymol. 218, 340–356] was used to synthesize the remaining segments of the full-length cDNA. The complete cDNA contained 1671 nucleotides with an open reading frame coding for 469 amino acid residues. The amino acid sequence deduced from the cDNA sequence included the amino acid sequences of all 12 trypsin digest fragments derived from the purified enzyme. Amino acid sequence analysis indicates that the guinea pig liver cytosolic β-glucosidase is a Family 1 β-glycosidase and that it is most closely related to mammalian lactase-phlorizin hydrolase. These results suggest that the cytosolic β-glucosidase and lactase-phlorizin hydrolase diverged from a common evolutionary precursor.

scholarly journals The C Terminus of Component C2II ofClostridium botulinum C2 Toxin Is Essential for Receptor Binding

2000 ◽  
Vol 68 (8) ◽  
pp. 4566-4573 ◽  
Author(s):  
Dagmar Blöcker ◽  
Holger Barth ◽  
Elke Maier ◽  
Roland Benz ◽  
Joseph T. Barbieri ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Receptor Binding ◽  
Sodium Dodecyl ◽  
Cytotoxic Action ◽  
Amino Acid Residues ◽  
C Terminus ◽  
Terminal Amino ◽  
Function Relationship ◽  
C2 Toxin

ABSTRACT The binary Clostridium botulinum C2 toxin consists of two separate proteins, the binding component C2II (80.5 kDa) and the actin-ADP-ribosylating enzyme component C2I (49.4 kDa). For its cytotoxic action, C2II binds to a cell membrane receptor and induces cell entry of C2I via receptor-mediated endocytosis. Here we studied the structure-function relationship of C2II by constructing truncated C2II proteins and producing polyclonal antisera against selective regions of C2II. An antibody raised against the C terminus (amino acids 592 to 721) of C2II inhibited binding of C2II to cells. The antibody prevented pore formation by C2II oligomers in artificial membranes but did not influence the properties of existing channels. To further define the region responsible for receptor binding, we constructed proteins with deletions in C2II; specifically, they lacked amino acid residues 592 to 721 and the 7 C-terminal amino acid residues. The truncated proteins still formed sodium dodecyl sulfate-stable oligomers but were unable to bind to cells. Our data indicate that the C terminus of C2II mediates binding of the protein to cells and that the 7 C-terminal amino acids are structurally important for receptor binding.

Structural requirements of pancreatic polypeptide receptor binding

1991 ◽  
Vol 261 (3) ◽  
pp. E319-E324
Author(s):  
R. L. Gingerich ◽  
J. O. Akpan ◽  
W. R. Gilbert ◽  
K. M. Leith ◽  
J. A. Hoffmann ◽  
...  
Keyword(s):  
Amino Acid ◽  
Methyl Ester ◽  
Receptor Binding ◽  
Pancreatic Polypeptide ◽  
Free Acid ◽  
Mammalian Species ◽  
Methionine Sulfoxide ◽  
Radioreceptor Assay ◽  
Amino Acid Residues ◽  
Structural Requirements

Pancreatic polypeptide (PP) receptors have been identified and characterized on the basolateral membranes (BLM) of canine intestinal mucosa. The present study was designed to ascertain the structural requirements of the PP molecule for binding to its receptor. A radioreceptor assay using purified BLM was employed to elucidate receptors specific to PPs of various mammalian species and to modified bovine PP (bPP) fragments. Receptor cross-reactivities (CR) to various PPs and bPP fragments were established. Results show that percent receptor CR by PPs of various species was as follows: bPP (100%) greater than human PP (68%) greater than porcine PP (50%) greater than canine PP (45%) greater than ovine PP (36%) greater than rat PP (3%). The fragments bPP-(1-15), bPP-(1-17), bPP-(1-26), bPP-(16-23), bPP-(18-30), bPP-(24-36), bPP-(27-35), and bPP-(31-36) at 500 nM did not significantly displace tracer from receptor (less than 0.1% CR). Des-COOH-terminal tyrosinamide [bPP-(1-35)] produced less than 0.1% CR. Oxidation of bPP methionine-30 residue to methionine sulfoxide decreased displacement to 67%. Modification of native amidated tyrosinamide to the free acid abolished receptor binding, whereas esterification to the methyl ester of COOH-terminal tyrosine restored binding to 60%. Additionally, percent CR decreased progressively as amino acid residues were deleted from the NH2-terminal region. We conclude that the molecular homologue of PP primary structure is necessary for full receptor binding. Both the NH2- and COOH-terminal residues are required for recognition, and the COOH-terminal tyrosinamide must be intact for PP binding to its receptor.

Synthesis and conformational studies of 3,4-di-O-acylated furanoid sugar amino acid-containing analogs of the receptor binding inhibitor of vasoactive intestinal peptide

2007 ◽  
Vol 48 (39) ◽  
pp. 6945-6950 ◽  
Author(s):  
T.K. Chakraborty ◽  
S. Uday Kumar ◽  
B. Krishna Mohan ◽  
G. Dattatreya Sarma ◽  
M. Udaya Kiran ◽  
...  
Keyword(s):  
Amino Acid ◽  
Vasoactive Intestinal Peptide ◽  
Receptor Binding ◽  
Conformational Studies
Sign in / Sign up

Export Citation Format

Share Document