Characterization of the Murine Platelet IIb Gene and Encoded cDNA

Blood ◽  
1999 ◽  
Vol 94 (11) ◽  
pp. 3947-3950
Author(s):  
Michael A. Thornton ◽  
Mortimer Poncz
Keyword(s):  
Amino Acids ◽  
Comparative Analysis ◽  
Platelet Aggregation ◽  
Ligand Binding ◽  
Human Gene ◽  
Cell System ◽  
Biological Studies ◽  
A Cell

The IIb/β3 receptor is central to platelet aggregation. Biological studies of this receptor have been limited by the inability to reproduce IIb/β3 function in a cell system. Increasingly, efforts are being directed at studies of this receptor in mice models. The structure of murine (m) β3 has been reported. We now have sequenced the mIIb gene and found that it has the same size and organization as the human gene. The exon/intron borders are reported here, as are the distances between exons. mIIb protein is 1,033 amino acids (aa), 7 and 5 aa shorter than human (h) and rodent (r) IIb, respectively, with 79% and 90% homology, respectively. As part of the comparative analysis of the 3 known IIb chains included in this report, we found that a particular region of the IIb N-terminal β-propeller is highly conserved and speculate that it directly participates in ligand binding.

Characterization of the Murine Platelet IIb Gene and Encoded cDNA

Blood ◽  
1999 ◽  
Vol 94 (11) ◽  
pp. 3947-3950 ◽  
Author(s):  
Michael A. Thornton ◽  
Mortimer Poncz
Keyword(s):  
Amino Acids ◽  
Comparative Analysis ◽  
Platelet Aggregation ◽  
Ligand Binding ◽  
Human Gene ◽  
Cell System ◽  
Biological Studies ◽  
A Cell

Abstract The IIb/β3 receptor is central to platelet aggregation. Biological studies of this receptor have been limited by the inability to reproduce IIb/β3 function in a cell system. Increasingly, efforts are being directed at studies of this receptor in mice models. The structure of murine (m) β3 has been reported. We now have sequenced the mIIb gene and found that it has the same size and organization as the human gene. The exon/intron borders are reported here, as are the distances between exons. mIIb protein is 1,033 amino acids (aa), 7 and 5 aa shorter than human (h) and rodent (r) IIb, respectively, with 79% and 90% homology, respectively. As part of the comparative analysis of the 3 known IIb chains included in this report, we found that a particular region of the IIb N-terminal β-propeller is highly conserved and speculate that it directly participates in ligand binding.

Proteomic Characterization of Protein-Protein Interactions

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Veenstra TD ◽  
Keyword(s):  
Protein Interactions ◽  
Disease Diagnosis ◽  
Cell System ◽  
Protein Protein Interactions ◽  
Novel Technologies ◽  
Cell Functions ◽  
Single Protein ◽  
A Cell ◽  
Molecular Components

Identifying all the molecular components within a living cell is the first step into understanding how it functions. To further understand how a cell functions requires identifying the interactions that occur between these components. This fact is especially relevant for proteins. No protein within a human cell functions on its own without interacting with another biomolecule - usually another protein. While Protein-Protein Interactions (PPI) have historically been determined by examining a single protein per study, novel technologies developed over the past couple of decades are enabling high-throughput methods that aim to describe entire protein networks within cells. In this review, some of the technologies that have led to these developments are described along with applications of these techniques. Ultimately the goal of these technologies is to map out the entire circuitry of PPI within human cells to be able to predict the global consequences of perturbations to the cell system. This predictive capability will have major impacts on the future of both disease diagnosis and treatment.

Isolation and partial characterization of amphibian tyrosine oxidase polysomes

Development ◽  
1970 ◽  
Vol 24 (1) ◽  
pp. 109-118
Author(s):  
E. L. Triplett ◽  
R. Herzog ◽  
L. P. Russell
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Enzymic Activity ◽  
Antigenic Determinants ◽  
Free System ◽  
A Cell ◽  
Generating System ◽  
Soluble Components

A population of polysomes isolated from frogskinis capable of supporting protein synthesis in a cell-free system containing an energy generating system, ‘soluble components’, and amino acids. These polysomes catalyse the oxidation of DOPA after gentle trypsinization, and they also have antigenic determinants attributable to tyrosine oxidase. Skin polysomes sedimented in 10–30 % sucrose gradients contain tyrosine oxidase peaks of enzymic activity at the bottom and top of the tube and in the 250 S regions. A peak of tyrosine oxidase antigenic acitvity is found in the 250–350S region of the gradient. Polysomes resolved on the gradient retain the ability to support protein synthesis in a cellfree system. All 250–350S particles capable of supporting the incorporation of [14C]amino acid into tyrosine oxidase are precipitable with tyrosine oxidase antibodies. It is probable that 250–350S tyrosine oxidase antibody precipitates contain only polysomes for this protein.

Characterization of estrogen receptor βb in sea bream (Sparus auratus): Phylogeny, ligand-binding, and comparative analysis of expression

2006 ◽  
Vol 145 (2) ◽  
pp. 197-207 ◽  
Author(s):  
Patrícia I.S. Pinto ◽  
Ana Lúcia Passos ◽  
Rute S. Martins, Deborah M. Power ◽  
Adelino V.M. Canário
Keyword(s):  
Estrogen Receptor ◽  
Comparative Analysis ◽  
Ligand Binding ◽  
Sea Bream ◽  
Sparus Auratus

scholarly journals Characterization of fundamental networks

2019 ◽  
Vol 150 (1) ◽  
pp. 453-474
Author(s):  
Manuela A. D. Aguiar ◽  
Ana P. S. Dias ◽  
Pedro Soares
Keyword(s):  
Network Architecture ◽  
Cell System ◽  
Hidden Symmetries ◽  
Network Construction ◽  
Cell Systems ◽  
Cell Network ◽  
A Cell ◽  
Network Synchrony ◽  
Coupled Cell Systems

AbstractIn the framework of coupled cell systems, a coupled cell network describes graphically the dynamical dependencies between individual dynamical systems, the cells. The fundamental network of a network reveals the hidden symmetries of that network. Subspaces defined by equalities of coordinates which are flow-invariant for any coupled cell system consistent with a network structure are called the network synchrony subspaces. Moreover, for every synchrony subspace, each network admissible system restricted to that subspace is a dynamical system consistent with a smaller network called a quotient network. We characterize networks such that: the network is a subnetwork of its fundamental network, and the network is a fundamental network. Moreover, we prove that the fundamental network construction preserves the quotient relation and it transforms the subnetwork relation into the quotient relation. The size of cycles in a network and the distance of a cell to a cycle are two important properties concerning the description of the network architecture. In this paper, we relate these two architectural properties in a network and its fundamental network.

scholarly journals Molecular characterization of the Agrobacterium tumefaciens DNA transfer protein VirB6

Microbiology ◽  
2005 ◽  
Vol 151 (11) ◽  
pp. 3483-3492 ◽  
Author(s):  
Paul K. Judd ◽  
David Mahli ◽  
Anath Das
Keyword(s):  
Amino Acids ◽  
Agrobacterium Tumefaciens ◽  
Transmembrane Domain ◽  
Ti Plasmid ◽  
Dna Transfer ◽  
Single Amino Acid ◽  
Dependent Manner ◽  
Cell Pole ◽  
A Cell

The VirB proteins of Agrobacterium tumefaciens assemble a T-pilus and a type IV secretion (T4S) apparatus for the transfer of DNA and proteins to plant cells. VirB6 is essential for DNA transfer and is a polytopic integral membrane protein with at least four membrane-spanning domains. VirB6 is postulated to function in T-pilus biogenesis and to be a component of the T4S apparatus. To identify amino acids required for VirB6 function, random mutations were introduced into virB6, and mutants that failed to complement a deletion in virB6 in tumour formation assays were isolated. Twenty-one non-functional mutants were identified, eleven of which had a point mutation that led to a substitution in a single amino acid. Characterization of the mutants indicated that the N-terminal large periplasmic domain and the transmembrane domain TM3 are required for VirB6 function. TM3 has an unusual sequence feature in that it is rich in bulky hydrophobic amino acids. This feature is found conserved in the VirB6 family of proteins. Studies on the effect of VirB6 on other VirB proteins showed that the octopine Ti-plasmid VirB6, unlike its nopaline Ti-plasmid counterpart, does not affect accumulation of VirB3 and VirB5, but has a strong negative effect on the accumulation of the VirB7-VirB7 dimer. Using indirect immunofluorescence microscopy the authors recently demonstrated that VirB6 localizes to a cell pole in a VirB-dependent manner. Mutations identified in the present study did not affect polar localization of the protein or the formation of the VirB7-VirB7 dimer. A VirB6-GFP fusion that contained the entire VirB6 ORF did not localize to a cell pole in either the presence or the absence of the other VirB proteins. IMF studies using dual labelling demonstrated that VirB6 colocalizes with VirB3 and VirB9, and not with VirB4, VirB5 and VirB11. These results support the conclusion that VirB6 is a structural component of the T4S apparatus.

Cloning and Characterization of Adinbitor, a Novel Disintegrin from the Snake Venom of Agkistrodon halys brevicaudus stejneger

2004 ◽  
Vol 36 (6) ◽  
pp. 425-429 ◽  
Author(s):  
Ji-Hong Wang ◽  
Yu Wu ◽  
Feng Ren ◽  
Li Lü ◽  
Bao-Chang Zhao
Keyword(s):  
Amino Acids ◽  
Platelet Aggregation ◽  
Human Platelet ◽  
Venom Gland ◽  
Rt Pcr ◽  
E Coli ◽  
Human Platelet Aggregation

Abstract Adinbitor was cloned from Agkistrodon halys brevicaudus stejneger and characterized as a novel disintegrin. In this study, total RNA was extracted from venom gland and used in RT-PCR to generate a cDNA which is 219 bp long. The sequence encoded a polypeptide composed of 73 amino acids, including 12 cysteines, an RGD motif, and the signature motif of disintergrin. Recombinant Adinbitor (rAdinbitor) was expressed in E. coli and purified by using the His·Bind affinity chromatography. The IC50 for inhibiting human platelet aggregation and bFGF-induced proliferation of ECV304 cells was 6 μM and 0.89 μM respectively. Furthermore, Adinbitor significantly inhibited angiogenesis both in vivo and in vitro. Taken together, these results suggested that Adinbitor had typical functions of disintegrins.

Functional Characterization of PM6/13, a β3-specific (GPIIIa/CD61) Monoclonal Antibody that Shows Preferential Inhibition of Fibrinogen Binding over Fibronectin Binding to Activated Human Platelets

1998 ◽  
Vol 79 (01) ◽  
pp. 177-185 ◽  
Author(s):  
Ashia Siddiqua ◽  
Michael Wilkinson ◽  
Vijay Kakkar ◽  
Yatin Patel ◽  
Salman Rahman ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Platelet Aggregation ◽  
Platelet Rich Plasma ◽  
Functional Characterization ◽  
Human Platelets ◽  
Western Blots ◽  
Activated Platelets ◽  
Reducing Conditions ◽  
Fibronectin Binding

SummaryWe report the characterization of a monoclonal antibody (MAb) PM6/13 which recognises glycoprotein IIIa (GPIIIa) on platelet membranes and in functional studies inhibits platelet aggregation induced by all agonists examined. In platelet-rich plasma, inhibition of aggregation induced by ADP or low concentrations of collagen was accompanied by inhibition of 5-hydroxytryptamine secretion. EC50 values were 10 and 9 [H9262]g/ml antibody against ADP and collagen induced responses respectively. In washed platelets treated with the cyclooxygenase inhibitor, indomethacin, PM6/13 inhibited platelet aggregation induced by thrombin (0.2 U/ml), collagen (10 [H9262]g/ml) and U46619 (3 [H9262]M) with EC50 = 4, 8 and 4 [H9262]g/ml respectively, without affecting [14C]5-hydroxytryptamine secretion or [3H]arachidonate release in appropriately labelled cells. Studies in Fura 2-labelled platelets revealed that elevation of intracellular calcium by ADP, thrombin or U46619 was unaffected by PM6/13 suggesting that the epitope recognised by the antibody did not influence Ca2+ regulation. In agreement with the results from the platelet aggregation studies, PM6/13 was found to potently inhibit binding of 125I-fibrinogen to ADP activated platelets. Binding of this ligand was also inhibited by two other MAbs tested, namely SZ-21 (also to GPIIIa) and PM6/248 (to the GPIIb-IIIa complex). However when tested against binding of 125I-fibronectin to thrombin stimulated platelets, PM6/13 was ineffective in contrast with SZ-21 and PM6/248, that were both potent inhibitors. This suggested that the epitopes recognised by PM6/13 and SZ-21 on GPIIIa were distinct. Studies employing proteolytic dissection of 125I-labelled GPIIIa by trypsin followed by immunoprecipitation with PM6/13 and analysis by SDS-PAGE, revealed the presence of four fragments at 70, 55, 30 and 28 kDa. PM6/13 did not recognize any protein bands on Western blots performed under reducing conditions. However Western blotting analysis with PM6/13 under non-reducing conditions revealed strong detection of the parent GP IIIa molecule, of trypsin treated samples revealed recognition of an 80 kDa fragment at 1 min, faint recognition of a 60 kDa fragment at 60 min and no recognition of any product at 18 h treatment. Under similar conditions, SZ-21 recognized fragments at 80, 75 and 55 kDa with the 55kDa species persisting even after 18 h trypsin treatment. These studies confirm the epitopes recognised by PM6/13 and SZ-21 to be distinct and that PM6/13 represents a useful tool to differentiate the characteristics of fibrinogen and fibronectin binding to the GPIIb-IIIa complex on activated platelets.

Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple

2018 ◽  
Vol 39 (4) ◽  
pp. 474-482
Author(s):  
Hoang Thi Le Thuong ◽  
Nguyen Quang Hao ◽  
Tran Thi Thuy
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Low Ph ◽  
Yeast Strains ◽  
Biological Studies ◽  
Taxonomic Characterization

Eight yeast strains (denoted as D1 to D8) were isolated from samples of natural fermented pineapple. Strain D8 showed highest alcoholic production at low pH and special aroma of pineapple has been chosen for further study. Taxonomic characterization of strain D8 using morphological, biochemical and molecular biological studies confirmed that strain D8  belong to Saccharomycetaceae family, Saccharomycetales order and Saccharomyces cerevisiae species. Therefore, we named this strain as Saccharomyces cerevisiae D8 for further study on Brandy production from pineapple. Citation: Hoang Thi Le Thuong, Nguyen Quang Hao, Tran Thi Thuy, 2017. Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple. Tap chi Sinh hoc, 39(4): 474- 482. DOI: 10.15625/0866-7160/v39n4.10864.*Corresponding author: [email protected] Received 5 December 2016, accepted 12 August 2017

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