scholarly journals Covalent cross-linking of the bovine somatotropin dimer. Effects on growth-promoting, receptor-binding and immunological activities and preliminary characterization of the self-association

1983 ◽  
Vol 209 (1) ◽  
pp. 107-115 ◽  
Author(s):  
H N Fernández ◽  
J M Delfino
Keyword(s):  
Critical Role ◽  
The Self ◽  
Bovine Somatotropin ◽  
Cross Linking ◽  
British Library ◽  
Dimethyl Suberimidate ◽  
Growth Promoting ◽  
Self Association

Bovine somatotropin, at pH 8.5 in 0.02 M-Bicine [NN-bis-(2-hydroxyethyl)glycine]/0.09M-NaCl, showed by frontal analysis the characteristics of a rapid monomer-dimer equilibrium whose dissociation constant was estimated to be 6.6×10(-6)M. Reaction of the hormone with dimethyl suberimidate lead to covalent cross-linking of the dimeric species. Under the conditions chosen (0.4 mg of bifunctional imidate and 1 mg of protein/ml at room temperature for 1 h) the cross-linked dimers accounted for 26% of the total protein, and these were isolated by molecular sieving in 0.29M-NH3/0.12M-NaCl. Covalent stabilization greatly diminished the growth-promoting activity and the ability to interact with somatogenic sites in both rat liver in vivo and rabbit liver microsomal fractions. Evidence indicating a non-critical role for amino groups involved in the covalent cross-linking was provided by a nearly equivalent derivative obtained after reaction with 3,3′-dithiobispropionimidate, which had substantial hormonal activity upon cleavage of the disulphide links. Conversely, immunological reactivity as demonstrated by radioimmunoassay was not affected by cross-linking. Details of the least-squares procedure employed to evaluate the self-association equilibrium constant has been deposited as Supplement SUP 50115 (7 pages) with the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies may be obtained on the terms indicated in Biochem. J. (1981) 193,5.

scholarly journals A biomimetic platelet based on assembling peptides initiates artificial coagulation

2020 ◽  
Vol 6 (22) ◽  
pp. eaaz4107
Author(s):  
Pei-Pei Yang ◽  
Kuo Zhang ◽  
Ping-Ping He ◽  
Yu Fan ◽  
Xuejiao J. Gao ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Binding Sites ◽  
Therapeutic Strategy ◽  
Critical Role ◽  
The Self ◽  
Membrane Glycoprotein ◽  
Tumor Site ◽  
Self Assembling ◽  
Receptor Interactions

Platelets play a critical role in the regulation of coagulation, one of the essential processes in life, attracting great attention. However, mimicking platelets for in vivo artificial coagulation is still a great challenge due to the complexity of the process. Here, we design platelet-like nanoparticles (pNPs) based on self-assembled peptides that initiate coagulation and form clots in blood vessels. The pNPs first bind specifically to a membrane glycoprotein (i.e., CD105) overexpressed on angiogenetic endothelial cells in the tumor site and simultaneously transform into activated platelet-like nanofibers (apNFs) through ligand-receptor interactions. Next, the apNFs expose more binding sites and recruit and activate additional pNPs, forming artificial clots in both phantom and animal models. The pNPs are proven to be safe in mice without systemic coagulation. The self-assembling peptides mimic platelets and achieve artificial coagulation in vivo, thus providing a promising therapeutic strategy for tumors.

Structural Characterization of a Biogenic Secretion Extracted from the Tendon or Ligament in Rabbits for Artificial Ligament Formation

2021 ◽  
Vol 1016 ◽  
pp. 786-791
Author(s):  
Toru Kuzumaki ◽  
Tatsuya Yamaguchi ◽  
Kengo Shimozaki ◽  
Junsuke Nakase ◽  
Kojun Torigoe
Keyword(s):  
Achilles Tendon ◽  
Collagen Fiber ◽  
Cross Linking ◽  
Fiber Structure ◽  
Collateral Ligament ◽  
Artificial Ligament ◽  
Model Method ◽  
Basic Investigation

Thus far, our research group has conducted a basic investigation for the development of an artificial ligament, which was performed by utilizing a biogenic secretion that was derived from the Achilles tendon in mice; this was achieved using the film model method. In this study, an attempt has been made to derive a biogenic secretion from the Achilles tendon (tendon gel) and the medial collateral ligament (ligament gel) in rabbits. Subsequently, a discussion was carried out on the possibility of forming a ligament-like structure that was based on the structural, mechanical, and spectroscopic investigations. The tendon gel was successfully formed from a parent tendon that was preserved in vivo for 3, 5, 10, and 15 d. Further, an aligned collagen fiber emerged in the tendon gel, which was subjected to tension on every preservation date. Further, the mechanical behavior of the tendon gel specimens was classified in two groups. The values of the Young's modulus of the specimens preserved for 10 and 15 d were higher than those of the specimens preserved for 3 and 5 d. Within the range of this experimental condition, the aligned collagen fiber structure was formed by applying a tension of approximately greater than 0.05 N. Conversely, only a 10-d preservation period yielded a sufficient amount of ligament gel for the experiment. Notably, the volume of ligament gel was less than that of the tendon gel. In the ligament gel specimen without the synovial membrane, the collagen fiber structure was formed by applying a tension, which was similar to that experienced by the tendon gel specimen. However, the cross-linking and growth of collagen fibers in the ligament gel samples were insignificant as compared with those of the tendon gel samples.

scholarly journals Identification of a Kinase-Active CheA Conformation in Escherichia coli Chemoreceptor Signaling Complexes

2019 ◽  
Vol 201 (23) ◽  
Author(s):  
Germán E. Piñas ◽  
John S. Parkinson
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Kinase Activity ◽  
Critical Role ◽  
Active State ◽  
Dynamics Simulation ◽  
Cross Linking ◽  
Atp Binding ◽  
Content Type

ABSTRACT Escherichia coli chemotaxis relies on control of the autophosphorylation activity of the histidine kinase CheA by transmembrane chemoreceptors. Core signaling units contain two receptor trimers of dimers, one CheA homodimer, and two monomeric CheW proteins that couple CheA activity to receptor control. Core signaling units appear to operate as two-state devices, with distinct kinase-on and kinase-off CheA output states whose structural nature is poorly understood. A recent all-atom molecular dynamic simulation of a receptor core unit revealed two alternative conformations, “dipped” and “undipped,” for the ATP-binding CheA.P4 domain that could be related to kinase activity states. To explore possible signaling roles for the dipped CheA.P4 conformation, we created CheA mutants with amino acid replacements at residues (R265, E368, and D372) implicated in promoting the dipped conformation and examined their signaling consequences with in vivo Förster resonance energy transfer (FRET)-based kinase assays. We used cysteine-directed in vivo cross-linking reporters for the dipped and undipped conformations to assess mutant proteins for these distinct CheA.P4 domain configurations. Phenotypic suppression analyses revealed functional interactions among the conformation-controlling residues. We found that structural interactions between R265, located at the N terminus of the CheA.P3 dimerization domain, and E368/D372 in the CheA.P4 domain played a critical role in stabilizing the dipped conformation and in producing kinase-on output. Charge reversal replacements at any of these residues abrogated the dipped cross-linking signal, CheA kinase activity, and chemotactic ability. We conclude that the dipped conformation of the CheA.P4 domain is critical to the kinase-active state in core signaling units. IMPORTANCE Regulation of CheA kinase in chemoreceptor arrays is critical for Escherichia coli chemotaxis. However, to date, little is known about the CheA conformations that lead to the kinase-on or kinase-off states. Here, we explore the signaling roles of a distinct conformation of the ATP-binding CheA.P4 domain identified by all-atom molecular dynamics simulation. Amino acid replacements at residues predicted to stabilize the so-called “dipped” CheA.P4 conformation abolished the kinase activity of CheA and its ability to support chemotaxis. Our findings indicate that the dipped conformation of the CheA.P4 domain is critical for reaching the kinase-active state in chemoreceptor signaling arrays.

In Vitro and In Vivo Characterization of Plant Growth Promoting Bacillus Strains Isolated from Extreme Environments of Eastern Algeria

2013 ◽  
Vol 172 (4) ◽  
pp. 1735-1746 ◽  
Author(s):  
Asma Ait-Kaki ◽  
Noreddine Kacem-Chaouche ◽  
Marc Ongena ◽  
Mounira Kara-Ali ◽  
Laid Dehimat ◽  
...  
Keyword(s):  
Plant Growth ◽  
Extreme Environments ◽  
Plant Growth Promoting ◽  
Growth Promoting ◽  
Eastern Algeria ◽  
In Vivo Characterization ◽  
Bacillus Strains

scholarly journals A new glycation product ‘norpronyl-lysine,’ and direct characterization of cross linking and other glycation adducts: NMR of model compounds and collagen

2014 ◽  
Vol 34 (2) ◽  
Author(s):  
Peter T. B. Bullock ◽  
David G. Reid ◽  
W. Ying Chow ◽  
Wendy P. W. Lau ◽  
Melinda J. Duer
Keyword(s):  
Model Systems ◽  
Cross Linking ◽  
Advanced Glycation ◽  
Model Compounds ◽  
Product Characterization ◽  
Glycation Product ◽  
Cross Links

NMR reveals numerous early and advanced glycation products, including a newly recognized ‘norpronyl-lysine,’ and cross links in solution, intact collagen and model systems. Solid state methods are directly applicable to in vitro and in vivo glycation pathway and product characterization.

scholarly journals A Ral GAP complex links PI 3-kinase/Akt signaling to RalA activation in insulin action

2011 ◽  
Vol 22 (1) ◽  
pp. 141-152 ◽  
Author(s):  
Xiao-Wei Chen ◽  
Dara Leto ◽  
Tingting Xiong ◽  
Genggeng Yu ◽  
Alan Cheng ◽  
...  
Keyword(s):  
Glucose Transporter ◽  
Critical Role ◽  
Inhibitory Effect ◽  
Small Gtpase ◽  
Regulatory Subunit ◽  
Hydrolysis Of ◽  
Identification And Characterization

Insulin stimulates glucose transport in muscle  and adipose tissue by translocation of glucose transporter 4 (GLUT4) to the plasma membrane. We previously reported that activation of the small GTPase RalA downstream of PI 3-kinase plays a critical role in this process by mobilizing the exocyst complex for GLUT4 vesicle targeting in adipocytes. Here we report the identification and characterization of a Ral GAP complex (RGC) that mediates the activation of RalA downstream of the PI 3-kinase/Akt pathway. The complex is composed of an RGC1 regulatory subunit and an RGC2 catalytic subunit (previously identified as AS250) that directly stimulates the guanosine triphosphate hydrolysis of RalA. Knockdown of RGC proteins leads to increased RalA activity and glucose uptake in adipocytes. Insulin inhibits the GAP complex through Akt2-catalyzed phosphorylation of RGC2 in vitro and in vivo, while activated Akt relieves the inhibitory effect of RGC proteins on RalA activity. The RGC complex thus connects PI 3-kinase/Akt activity to the transport machineries responsible for GLUT4 translocation.

scholarly journals Full in vivo characterization of carbonate chemistry at the site of calcification in corals

2019 ◽  
Vol 5 (1) ◽  
pp. eaau7447 ◽  
Author(s):  
Duygu S. Sevilgen ◽  
Alexander A. Venn ◽  
Marian Y. Hu ◽  
Eric Tambutté ◽  
Dirk de Beer ◽  
...  
Keyword(s):  
Critical Role ◽  
Carbonate Chemistry ◽  
Saturation State ◽  
Stylophora Pistillata ◽  
Coral Calcification ◽  
Carbon Concentrating Mechanisms ◽  
In Vivo Characterization

Reef-building corals form their calcium carbonate skeletons within an extracellular calcifying medium (ECM). Despite the critical role of the ECM in coral calcification, ECM carbonate chemistry is poorly constrained in vivo, and full ECM carbonate chemistry has never been characterized based solely on direct in vivo measurements. Here, we measure pHECMin the growing edge ofStylophora pistillataby simultaneously using microsensors and the fluorescent dye SNARF-1, showing that, when measured at the same time and place, the results agree. We then conduct microscope-guided microsensor measurements of pH, [Ca2+], and [CO32−] in the ECM and, from this, determine [DIC]ECMand aragonite saturation state (Ωarag), showing that all parameters are elevated with respect to the surrounding seawater. Our study provides the most complete in vivo characterization of ECM carbonate chemistry parameters in a coral species to date, pointing to the key role of calcium- and carbon-concentrating mechanisms in coral calcification.

In Vitro and In Vivo Characterization of Scleral Implant of Indomethacin: Role of Plasticizer and Cross-Linking Time

Drug Delivery ◽  
2003 ◽  
Vol 10 (4) ◽  
pp. 269-275 ◽  
Author(s):  
M. Thilek Kumar ◽  
C. Rajeswari ◽  
J. Balasubramaniam ◽  
J. K. Pandit ◽  
S. Kant
Keyword(s):  
Cross Linking ◽  
In Vivo Characterization
Sign in / Sign up

Export Citation Format

Share Document