scholarly journals Three Dimensional Structure of Blood Coagulation Factor Xa.

1999 ◽  
Vol 10 (2/3) ◽  
pp. 181-188 ◽  
Author(s):  
Kenji KAMATA ◽  
Toshiyuki MIYATA
Keyword(s):  
Blood Coagulation ◽  
Three Dimensional ◽  
Factor Xa ◽  
Coagulation Factor ◽  
Dimensional Structure ◽  
Three Dimensional Structure

Three-dimensional structure of the apo form of the N-terminal EGF-like module of blood coagulation factor X as determined by NMR spectroscopy and simulated folding

Biochemistry ◽  
1992 ◽  
Vol 31 (26) ◽  
pp. 5974-5983 ◽  
Author(s):  
Magnus Ullner ◽  
Maria Selander ◽  
Egon Persson ◽  
Johan Stenflo ◽  
Torbjoern Drakenberg ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Blood Coagulation ◽  
Three Dimensional ◽  
Coagulation Factor ◽  
Dimensional Structure ◽  
Factor X ◽  
Three Dimensional Structure

scholarly journals Crystal Structure of Isoform CBd of the Basic Phospholipase A2 Subunit of Crotoxin: Description of the Structural Framework of CB for Interaction with Protein Targets

Molecules ◽  
2020 ◽  
Vol 25 (22) ◽  
pp. 5290
Author(s):  
Dorota Nemecz ◽  
Maciej Ostrowski ◽  
Marc Ravatin ◽  
Frederick Saul ◽  
Grazyna Faure
Keyword(s):  
Cystic Fibrosis ◽  
Phospholipase A2 ◽  
Three Dimensional ◽  
Factor Xa ◽  
Coagulation Factor ◽  
Presynaptic Receptors ◽  
Dimensional Structure ◽  
Protein Targets ◽  
Structural Framework ◽  
Crotalus Durissus

Crotoxin, from the venom of the South American rattlesnake Crotalus durissus terrificus, is a potent heterodimeric presynaptic β-neurotoxin that exists in individual snake venom as a mixture of isoforms of a basic phospholipase A2 (PLA2) subunit (CBa2, CBb, CBc, and CBd) and acidic subunit (CA1–4). Specific natural mutations in CB isoforms are implicated in functional differences between crotoxin isoforms. The three-dimensional structure of two individual CB isoforms (CBa2, CBc), and one isoform in a crotoxin (CA2CBb) complex, have been previously reported. This study concerns CBd, which by interaction with various protein targets exhibits many physiological or pharmacological functions. It binds with high affinity to presynaptic receptors showing neurotoxicity, but also interacts with human coagulation factor Xa (hFXa), exhibiting anticoagulant effect, and acts as a positive allosteric modulator and corrector of mutated chloride channel, cystic fibrosis transmembrane conductance regulator (CFTR), implicated in cystic fibrosis. Thus, CBd represents a novel family of agents that have potential in identifying new drug leads related to anticoagulant and anti-cystic fibrosis function. We determined here the X-ray structure of CBd and compare it with the three other natural isoforms of CB. The structural role of specific amino acid variations between CB isoforms are analyzed and the structural framework of CB for interaction with protein targets is described.

Cellular Consequences of the Initiation of Blood Coagulation

1999 ◽  
Vol 82 (08) ◽  
pp. 183-192 ◽  
Author(s):  
Eric Camerer ◽  
John-Arne Røttingen ◽  
Merete Thune Wiiger ◽  
Elisabet Gjernes ◽  
Hans Prydz
Keyword(s):  
Endothelial Cells ◽  
Blood Coagulation ◽  
Intracellular Signaling ◽  
Blood Clotting ◽  
Factor Viia ◽  
Three Dimensional ◽  
Factor Xa ◽  
Factor Vii ◽  
Dimensional Structure ◽  
Major Player

IntroductionThis paper reviews some of the cell biological aspects of the consequences of blood clotting initiation. These intracellular events occur in cells carrying tissue factor (TF) when its ligand, factor VIIa, is bound to the receptor-like TF surface molecules. The intracellular signaling generated by this ligand/receptor binding and some of its consequences are described and parallel experiments with factor Xa are discussed.The role of TF as a major player in the initiation of blood coagulation has been known since the last century1,2 and is now characterized in molecular detail. Research on TF, for a long period and for obvious reasons, concentrated on its essential role as a cofactor in this process. Its importance in the development of clinical thrombosis, be it venous or arterial, has been appreciated since it was discovered that monocytes and macrophages3 and endothelial cells,4 under certain conditions, could be induced to synthesize TF. This contributed to answering the previously unresolved question about how TF got into contact with the flowing blood in the absence of any trauma. We later demonstrated that the TF induction process, in many cases, is subject to down-regulation by cAMP5,6 and that Ca2+ influx can induce the synthesis,5,6 along with a large number of other compounds.7 We also showed that protein kinase C was a mediator in at least some of these inducing pathways.8 The purification of TF in 19739 showed that TF was an integral membrane protein. By 1977 it was clear that TF likely participated in functions other than blood clotting.10 The cloning of the gene for TF11-14 suggested that, structurally, TF was a member of the Class II cytokine receptor family.15 To fulfil the criteria for being a true receptor, it also needed a specific and high-affinity ligand, which it has in factor VII. Also, to be classified as a true receptor, ligand binding should generate an intracellular signal. In 1992, we presented the first report of such a signal in the form of Ca2+ peaks. These peaks were triggered by the addition of factor VIIa to endothelial cells carrying TF on their surface as a result of exposure to interleukin 1β. These signals were characterized further16,17 and were thought to render final proof for the function of the TF receptor.This review discusses our findings with respect to TF/factor VIIa-induced intracellular Ca2+-signaling and concludes that there is likely a two-component receptor. The more consequential question—whether this intracellular signaling leads to altered gene expression and to other phenotypic changes—is also raised. The establishment of knockout mice18–20 and efforts to solve the three-dimensional structure of this complex by x-ray diffraction21–24 are not reviewed extensively.

The Three-dimensional structure of frozen-hydrated nudaurelia capensis β virus

1987 ◽  
Vol 45 ◽  
pp. 650-651
Author(s):  
N. H. Olson ◽  
T. S. Baker ◽  
Wu Bo Mu ◽  
J. E. Johnson ◽  
D. A. Hendry
Keyword(s):  
South African ◽  
Rna Virus ◽  
Carbon Film ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Electron Dose ◽  
Total Electron ◽  
Three Dimensional Structure ◽  
Negative Stain ◽  
Dimensional Reconstruction

Nudaurelia capensis β virus (NβV) is an RNA virus of the South African Pine Emperor moth, Nudaurelia cytherea capensis (Lepidoptera: Saturniidae). The NβV capsid is a T = 4 icosahedron that contains 60T = 240 subunits of the coat protein (Mr = 61,000). A three-dimensional reconstruction of the NβV capsid was previously computed from visions embedded in negative stain suspended over holes in a carbon film. We have re-examined the three-dimensional structure of NβV, using cryo-microscopy to examine the native, unstained structure of the virion and to provide a initial phasing model for high-resolution x-ray crystallographic studiesNβV was purified and prepared for cryo-microscopy as described. Micrographs were recorded ∼1 - 2 μm underfocus at a magnification of 49,000X with a total electron dose of about 1800 e-/nm2.

Three Dimensional structure and organization of diploid chromosomes by optical section microscopy

1987 ◽  
Vol 45 ◽  
pp. 633-635
Author(s):  
David A. Agard ◽  
Yasushi Hiraoka ◽  
John W. Sedat
Keyword(s):  
Dimensional Space ◽  
Three Dimensional ◽  
Developmental State ◽  
Mitotic Cell ◽  
Biological Properties ◽  
Dimensional Structure ◽  
Specific Gene ◽  
Three Dimensional Structure ◽  
Complementary Techniques ◽  
Dynamic Structures

In an effort to understand the complex relationship between structure and biological function within the nucleus, we have embarked on a program to examine the three-dimensional structure and organization of Drosophila melanogaster embryonic chromosomes. Our overall goal is to determine how DNA and proteins are organized into complex and highly dynamic structures (chromosomes) and how these chromosomes are arranged in three dimensional space within the cell nucleus. Futher, we hope to be able to correlate structual data with such fundamental biological properties as stage in the mitotic cell cycle, developmental state and transcription at specific gene loci.Towards this end, we have been developing methodologies for the three-dimensional analysis of non-crystalline biological specimens using optical and electron microscopy. We feel that the combination of these two complementary techniques allows an unprecedented look at the structural organization of cellular components ranging in size from 100A to 100 microns.

Three-Dimensional Structure of Cloned T3 Connector Protein at 1.6nm Resolution

1990 ◽  
Vol 48 (1) ◽  
pp. 282-283
Author(s):  
José L. Carrascosa ◽  
José M. Valpuesta ◽  
Hisao Fujisawa
Keyword(s):  
Dna Sequences ◽  
Expression Vector ◽  
Three Dimensional ◽  
Deae Cellulose ◽  
Dna Packaging ◽  
Dimensional Structure ◽  
Two Dimensional ◽  
Freezing And Thawing ◽  
Three Dimensional Structure ◽  
Dimensional Reconstruction

The head to tail connector of bacteriophages plays a fundamental role in the assembly of viral heads and DNA packaging. In spite of the absence of sequence homology, the structure of connectors from different viruses (T4, Ø29, T3, P22, etc) share common morphological features, that are most clearly revealed in their three-dimensional structure. We have studied the three-dimensional reconstruction of the connector protein from phage T3 (gp 8) from tilted view of two dimensional crystals obtained from this protein after cloning and purification.DNA sequences including gene 8 from phage T3 were cloned, into Bam Hl-Eco Rl sites down stream of lambda promotor PL, in the expression vector pNT45 under the control of cI857. E R204 (pNT89) cells were incubated at 42°C for 2h, harvested and resuspended in 20 mM Tris HC1 (pH 7.4), 7mM 2 mercaptoethanol, ImM EDTA. The cells were lysed by freezing and thawing in the presence of lysozyme (lmg/ml) and ligthly sonicated. The low speed supernatant was precipitated by ammonium sulfate (60% saturated) and dissolved in the original buffer to be subjected to gel nitration through Sepharose 6B, followed by phosphocellulose colum (Pll) and DEAE cellulose colum (DE52). Purified gp8 appeared at 0.3M NaCl and formed crystals when its concentration increased above 1.5 mg/ml.

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