Cryoelectron microscopy of complexes of human rhinovirus with a monoclonal FAB and the viral cellular receptor

1992 ◽  
Vol 50 (1) ◽  
pp. 524-525
Author(s):  
Norman H. Olson ◽  
Thomas J. Smith ◽  
Prasanna R. Kolatkar ◽  
Marcos A. Oliveira ◽  
Roland R. Rueckert ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Antibody Fragment ◽  
Human Rhinovirus ◽  
Dimensional Structure ◽  
Intercellular Adhesion Molecule ◽  
Cellular Receptor ◽  
Intercellular Adhesion Molecule 1 ◽  
Amino Terminal ◽  
Functional Relationships ◽  
Neutralizing Monoclonal Antibody

Cryo-electron microscopy and image analysis techniques make it possible to study structural and functional relationships of macromolecular complexes that currently are not easily examined with crystallographic techniques. We have recorded images of frozen-hydrated human rhinovirus serotype-14 (HRV-14) complexed with a neutralizing, monoclonal, antibody fragment (Fab-17Ia; Fig. 1 A); and HRV-16 complexed with the amino-terminal, two-domain fragment (D1D2) of its cellular receptor (intercellular adhesion molecule-1, ICAM-1; Fig. 1B). Three-dimensional reconstructions (Figs. 2A,B) were calculated to ∽3nm resolution from 35 and 44 images of each complex, respectively. The HRV-14/Fab structure clearly identified the footprint of the Fab on the surface of the virion. The HRV-16/D1D2 reconstruction presents, for the first time, the three-dimensional structure of a complete virus complexed with its cellular receptor.

scholarly journals Interaction of Coxsackievirus A21 with Its Cellular Receptor, ICAM-1

2001 ◽  
Vol 75 (5) ◽  
pp. 2444-2451 ◽  
Author(s):  
Chuan Xiao ◽  
Carol M. Bator ◽  
Valorie D. Bowman ◽  
Elizabeth Rieder ◽  
Yongning He ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Adhesion Molecule ◽  
Intercellular Adhesion Molecule ◽  
Cellular Receptor ◽  
Intercellular Adhesion Molecule 1 ◽  
Common Region ◽  
Amino Terminal ◽  
Receptor Recognition ◽  
Coxsackievirus A21 ◽  
The Common

ABSTRACT Coxsackievirus A21 (CAV21), like human rhinoviruses (HRVs), is a causative agent of the common cold. It uses the same cellular receptor, intercellular adhesion molecule 1 (ICAM-1), as does the major group of HRVs; unlike HRVs, however, it is stable at acid pH. The cryoelectron microscopy (cryoEM) image reconstruction of CAV21 is consistent with the highly homologous crystal structure of poliovirus 1; like other enteroviruses and HRVs, CAV21 has a canyon-like depression around each of the 12 fivefold vertices. A cryoEM reconstruction of CAV21 complexed with ICAM-1 shows all five domains of the extracellular component of ICAM-1. The known atomic structure of the ICAM-1 amino-terminal domains D1 and D2 has been fitted into the cryoEM density of the complex. The site of ICAM-1 binding within the canyon of CAV21 overlaps the site of receptor recognition utilized by rhinoviruses and polioviruses. Interactions within this common region may be essential for triggering viral destabilization after attachment to susceptible cells.

scholarly journals Discrimination among Rhinovirus Serotypes for a Variant ICAM-1 Receptor Molecule

2004 ◽  
Vol 78 (18) ◽  
pp. 10034-10044 ◽  
Author(s):  
Chuan Xiao ◽  
Tobias J. Tuthill ◽  
Carol M. Bator Kelly ◽  
Lisa J. Challinor ◽  
Paul R. Chipman ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Salt Bridge ◽  
Human Rhinovirus ◽  
Intercellular Adhesion Molecule ◽  
Stable Complex ◽  
Intercellular Adhesion Molecule 1 ◽  
Binding Characteristics ◽  
X Ray Crystallography ◽  
Density Maps ◽  
Charge Interaction

ABSTRACT Intercellular adhesion molecule 1 (ICAM-1) is the cellular receptor for the major group of human rhinovirus serotypes, including human rhinovirus 14 (HRV14) and HRV16. A naturally occurring variant of ICAM-1, ICAM-1Kilifi, has altered binding characteristics with respect to different HRV serotypes. HRV14 binds to ICAM-1 only transiently at physiological temperatures but forms a stable complex with ICAM-1Kilifi. Conversely, HRV16 forms a stable complex with ICAM-1 but does not bind to ICAM-1Kilifi. The three-dimensional structures of HRV14 and HRV16, complexed with ICAM-1, and the structure of HRV14, complexed with ICAM-1Kilifi, have been determined by cryoelectron microscopy (cryoEM) image reconstruction to a resolution of approximately 10 Å. Structures determined by X-ray crystallography of both viruses and of ICAM-1 were fitted into the cryoEM density maps. The interfaces between the viruses and receptors contain extensive ionic networks. However, the interactions between the viruses and ICAM-1Kilifi contain one less salt bridge than between the viruses and ICAM-1. As HRV16 has fewer overall interactions with ICAM-1 than HRV14, the absence of this charge interaction has a greater impact on the binding of ICAM-1Kilifi to HRV16 than to HRV14.

Artificial Myocardium: Design Principles and Substratum

2001 ◽  
Vol 7 (S2) ◽  
pp. 138-139
Author(s):  
Michael Yost ◽  
Robert Price ◽  
David Simpson ◽  
Louis Terracio
Keyword(s):  
Gap Junctions ◽  
Three Dimensional ◽  
Surgical Techniques ◽  
Disease Process ◽  
The United States ◽  
Dimensional Structure ◽  
Muscular Tissue ◽  
Cardiac Muscle Cells ◽  
Functional Relationships ◽  
Electrical Pacing

Death and disability due to cardiovascular disease and congenital anomalies remains a significant health problem in the United States. Despite improvements in detection, patient management, surgery and preventative medicine, the quality of life for people who suffer from cardiovascular dysfunction has a major impact on our society. The intact heart is an elaborate three-dimensional structure that insures the orderly propagation of electrical signals coordinating the contraction and relaxation of the ventricular wall. Localized loss of muscular tissue as a result of congenital defect or disease process alters this structural arrangement and impairs overall cardiac function. Conventional surgical techniques cannot begin to adequately restore the subtle structural and functional relationships in the heart. The ability to construct a tissue-engineered prosthesis composed of cardiac muscle cells in a collagen-based scaffold may potentially offer a superior alternative to currently available surgical techniques.A tissue engineered myocardium must have the following components: First, it must develop and maintain the correct cellular phenotype as well as a functioning contractile apparatus of parallel myofibrils, Second it must be able to form gap junctions within itself and with the native tissue and these gap junctions must be competent at conducting electrical pacing as well as other biochemical signals, and Third, it must be capable of participating in and contributing to the rhythmic contraction of normal myocardium as well as accommodate the changes in contraction frequency ubiquitous in the cardiac environment

scholarly journals Optimization of the crystallizability of a single-chain antibody fragment

2014 ◽  
Vol 70 (12) ◽  
pp. 1701-1706 ◽  
Author(s):  
Jana Škerlová ◽  
Vlastimil Král ◽  
Milan Fábry ◽  
Juraj Sedláček ◽  
Václav Veverka ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Antibody Fragment ◽  
Binding Mode ◽  
Antibody Fragments ◽  
Dimensional Structure ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Diagnostic Potential ◽  
Storage Buffer ◽  
Single Chain Antibody Fragment

Single-chain variable antibody fragments (scFvs) are molecules with immense therapeutic and diagnostic potential. Knowledge of their three-dimensional structure is important for understanding their antigen-binding mode as well as for protein-engineering approaches such as antibody humanization. A major obstacle to the crystallization of single-chain variable antibody fragments is their relatively poor homogeneity caused by spontaneous oligomerization. A new approach to optimization of the crystallizability of single-chain variable antibody fragments is demonstrated using a representative single-chain variable fragment derived from the anti-CD3 antibody MEM-57. A Thermofluor-based assay was utilized to screen for optimal conditions for antibody-fragment stability and homogeneity. Such an optimization of the protein storage buffer led to a significantly improved ability of the scFv MEM-57 to yield crystals.

In vitro Inhibitory Activity of Soluble ICAM-1 for the Numbered Serotypes of Human Rhinovirus

1993 ◽  
Vol 4 (6) ◽  
pp. 323-327 ◽  
Author(s):  
C. E. Crump ◽  
E. Arruda ◽  
F. G. Hayden
Keyword(s):  
Human Rhinovirus ◽  
Intercellular Adhesion Molecule ◽  
Yield Reduction ◽  
Inhibitory Effects ◽  
Intercellular Adhesion Molecule 1 ◽  
Human Embryonic Lung ◽  
Group Type ◽  
Hel Cells ◽  
Soluble Intercellular Adhesion Molecule

We studied the antirhinovirus activity of soluble intercellular adhesion molecule-1 (sICAM-1) against all 100 numbered human rhinovirus (HRV) serotypes in human embryonic lung fibroblast (HEL) cells. sICAM-1 inhibited replication of 88 of 90 HRVs belonging to the major receptor group with 50% effective concentrations (EC50) ranging from 0.1 to 41.1 μg ml−1 (~0.002 to 0.8 μM). No inhibition at 100 μg ml−1 (~2 μM) was observed for HRV belonging to the minor receptor group, type 87, and two serotypes of the major receptor group (types 23 and 25). Yield reduction assays done with five HRV serotypes in human adenoid expiants found that sICAM-1 had concentration-dependent inhibitory effects that correlated closely with the activity observed in HEL cells ( r2 = 0.99, P < 0.001).

scholarly journals Enterovirus Capsid Interactions with Decay-Accelerating Factor Mediate Lytic Cell Infection

2004 ◽  
Vol 78 (3) ◽  
pp. 1431-1439 ◽  
Author(s):  
Nicole G. Newcombe ◽  
E. Susanne Johansson ◽  
Gough Au ◽  
A. Michael Lindberg ◽  
Richard D. Barry ◽  
...  
Keyword(s):  
Clinical Isolates ◽  
Intercellular Adhesion Molecule ◽  
Cross Linking ◽  
Target Cells ◽  
Cellular Receptor ◽  
Intercellular Adhesion Molecule 1 ◽  
Cell Infection ◽  
Decay Accelerating Factor ◽  
Receptor Usage ◽  
Cellular Attachment

ABSTRACT The cellular receptor usage of numerous human enteroviruses can differ significantly between low-cell-culture-passaged clinical isolates and highly laboratory-passaged prototype strains. The prototype strain of coxsackievirus A21 (CVA21) displays a dual-receptor specificity as determined with a receptor complex consisting of decay-accelerating factor (DAF) and intercellular adhesion molecule 1 (ICAM-1). In this study, the cellular receptor interactions of low-cell-passage CVA21 clinical isolates with respect to their interactions with cell surface-expressed DAF and ICAM-1 were compared to those of the CVA21 prototype (Kuykendall) strain. Dual-receptor usage of DAF and ICAM-1 by CVA21 clinical isolates was confirmed by cell transfection and radiolabeled binding assays. The cellular attachment of clinical and prototype CVA21 strains to cells that coexpressed DAF and ICAM-1 was not additive compared to the viral binding to cells expressing one or other receptor. In fact, the binding data suggest there is an inhibition of CVA21 cellular attachment in environments where high-level coexpression of both DAF and ICAM-1 occurs. Antibody cross-linking of DAF rendered cells susceptible to lytic infection by the CVA21 clinical isolates. In a novel finding, three clinical isolates could, to various degrees, infect and lyse DAF-expressing cells in the absence of DAF-antibody cross-linking and ICAM-1 expression. Sequence analysis of the P1 region of clinical and prototype virus genomes identified a number of coding changes that may contribute to the observed enhanced DAF usage phenotype of the clinical CVA21 isolates. None of the amino acid changes was located in the previously postulated ICAM-1 footprint, a receptor-binding environment that was conserved on the capsid surface of all CVA21 clinical isolates. Taken together, the data suggest that community-circulating strains of CVA21 can infect target cells expressing either ICAM-1 or DAF alone and that such interactions extend tissue tropism and impact directly on viral pathogenesis.

scholarly journals Molecular cloning of the rat analogue of human CD59: structural comparison with human CD59 and identification of a putative active site

1994 ◽  
Vol 304 (2) ◽  
pp. 595-601 ◽  
Author(s):  
N K Rushmere ◽  
R A Harrison ◽  
C W van den Berg ◽  
B P Morgan
Keyword(s):  
Molecular Cloning ◽  
Rat Kidney ◽  
Amino Acid Level ◽  
Three Dimensional ◽  
Glycosylation Site ◽  
Dimensional Structure ◽  
Flanking Sequence ◽  
Coding Region ◽  
Amino Terminal ◽  
Human Cd59

We have previously described the purification and partial characterization of the rat analogue of the human complement regulatory molecule CD59 [Hughes, Piddlesden, Williams, Harrison and Morgan (1992) Biochem. J. 284, 169-176]. We present here the molecular cloning and full sequence analysis of this molecule. A PCR-based approach utilizing primers designed from the amino-terminal protein sequence was used to isolate a full-length cDNA clone from a rat kidney cDNA library. This clone encoded a 92 bp 5′-flanking sequence, a 66 bp signal peptide and a 315 bp coding region containing putative glycosylation and GPI-anchor signals. The 3′ untranslated flanking region was approximately 1.1 kbp long and included the poly-A tail and a CATA repeating sequence. The coding region was 58% identical with the human cDNA at the nucleotide level and 44% identical at the amino acid level. Despite this relatively low overall sequence conservation, several highly conserved stretches were apparent, particularly in the N-terminal portion of the molecule, in the cysteine-rich region immediately preceding the site of glycolipid attachment and in the C-terminal peptide removed during glycolipid attachment. An N-glycosylation site was identified at Asn-16 and a putative glycosylphosphatidylinositol anchor addition site at Asn-79, indicating that the mature processed protein was two residues longer than human CD59. Comparison of the sequences of rat and human CD59, together with consideration of the published three-dimensional structure of human CD59 and functional data, implicates specific regions of the protein in interactions with C-8 and/or C-9.

scholarly journals Three-dimensional structure of the amino-terminal domain of syntaxin 6, a SNAP-25 C homolog

2002 ◽  
Vol 99 (14) ◽  
pp. 9184-9189 ◽  
Author(s):  
K. M. S. Misura ◽  
J. B. Bock ◽  
L. C. Gonzalez ◽  
R. H. Scheller ◽  
W. I. Weis
Keyword(s):  
Three Dimensional ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Amino Terminal Domain

scholarly journals Key Proteins Involved in Spheroid Formation and Angiogenesis in Endothelial Cells After Long-Term Exposure to Simulated Microgravity

2018 ◽  
Vol 45 (2) ◽  
pp. 429-445 ◽  
Author(s):  
Anita Dittrich ◽  
Daniela Grimm ◽  
Jayashree Sahana ◽  
Johann Bauer ◽  
Marcus Krüger ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Adhesion Molecule ◽  
Cell Structure ◽  
Three Dimensional ◽  
Cardiovascular Complications ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Multicellular Spheroids ◽  
Monocyte Chemoattractant Protein 1 ◽  
Molecular Approaches

Background/Aims: Cardiovascular complications are common in astronauts returning from a prolonged spaceflight. These health problems might be driven by complex modulations of gene expression and protein synthesis in endothelial cells (ECs). Studies on the influence of microgravity on phenotype, growth pattern and biological processes of ECs can help to understand these complications. Methods: We exposed ECs (EA.hy926) to a Random Positioning Machine (RPM). Proteins associated with cell structure, angiogenesis and endothelial dysfunction were investigated in distinct pools of multicellular spheroids (MCS), adherent cells (AD) and tubular structures (TS) formed after a 35-day RPM-exposure. Results: Combining morphological and molecular approaches, we found AD, MCS and TS with changes in the synthesis and release of proteins involved in three-dimensional growth. Fibronectin and monocyte chemoattractant protein-1 (MCP-1) mRNAs and protein contents were elevated along with an increased secretion of vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, MCP-1, intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), neutrophil gelatinase-associated lipocalin (NGAL) and regulated on activation, normal T cell expressed and secreted (RANTES) proteins in the culture supernatant as determined by multianalyte profiling technology. Together they form a network of interaction. Conclusions: These results show that a prolonged RPM-exposure of ECs induced TS and MCS formation. The factors VEGF, NGAL, IL-6, IL-8, MCP-1, VCAM-1, ICAM-1, fibronectin and RANTES seem to be affected when gravity is omitted.

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