Characterization of STAT Self-Association by Analytical Ultracentrifugation

2012 ◽  
pp. 203-224 ◽  
Author(s):  
Nikola Wenta ◽  
Uwe Vinkemeier
Keyword(s):  
Analytical Ultracentrifugation ◽  
Self Association

scholarly journals RNA Multimerisation in the DNA Packaging Motor of Bacteriophage φ29

2005 ◽  
Vol 6 (2) ◽  
pp. 127-134 ◽  
Author(s):  
Jonathan P. A. Wood ◽  
Stephanie A. Capaldi ◽  
Mark A. Robinson ◽  
Andrew J. Baron ◽  
Nicola J. Stonehouse
Keyword(s):  
Analytical Ultracentrifugation ◽  
Molecular Level ◽  
Viral Assembly ◽  
Partial Specific Volume ◽  
Wild Type ◽  
Rna Molecules ◽  
Dna Packaging Motor ◽  
Self Association

The use of bacteriophages as experimental tools allows the investigation of interactions between components at the molecular level that are often not possible in more complex virus systems. The bacteriophage φ29 acts as a molecular machine to package its own genomic DNA during viral assembly. Self-associating RNA molecules, called pRNA, have an essential role in the function of this machine. This paper reports the characterization of this self-association (which leads to multimerisation of wild-type and truncated variant pRNAs) by analytical ultracentrifugation (including determination of the partial specific volume of the pRNA), together with an investigation into the domains of the molecule important for multimerisation by the use of complementary DNA probes.

scholarly journals Characterization of the solution properties and conformation of pneumolysin, the membrane-damaging toxin of Streptococcus pneumoniae

1993 ◽  
Vol 296 (3) ◽  
pp. 671-674 ◽  
Author(s):  
P J Morgan ◽  
P G Varley ◽  
A J Rowe ◽  
P W Andrew ◽  
T J Mitchell
Keyword(s):  
Streptococcus Pneumoniae ◽  
Tertiary Structure ◽  
Analytical Ultracentrifugation ◽  
Sedimentation Coefficient ◽  
Spectroscopic Methods ◽  
Solution Conformation ◽  
Structure Information ◽  
Self Association ◽  
Secondary And Tertiary Structure

Pneumolysin is a membrane-damaging toxin produced by Streptococcus pneumoniae. In order to understand fully the mode of action of this toxin, it is necessary to have an appreciation of the size, self-association behaviour and solution conformation of pneumolysin. A combination of analytical ultracentrifugation methodologies has shown that pneumolysin lacks self-association behaviour in solution and has provided a weight-average M(r) (M omega) of 52,000 +/- 2000, which was in agreement with that derived from the amino acid sequence. By determining a sedimentation coefficient (S20,w0) of 3.35 +/- 0.10 S, it was possible to suggest a model for the gross solution conformation of pneumolysin monomers. Spectroscopic methods provide additional secondary and tertiary structure information.

scholarly journals Characterization of the Self Association ofAvian SarcomaVirus Integrase by Analytical Ultracentrifugation

1999 ◽  
Vol 274 (46) ◽  
pp. 32842-32846 ◽  
Author(s):  
Jacqueline Coleman ◽  
Steven Eaton ◽  
George Merkel ◽  
Anna Marie Skalka ◽  
Thomas Laue
Keyword(s):  
Analytical Ultracentrifugation ◽  
The Self ◽  
Self Association

scholarly journals Characterization of Native Reversible Self-Association of a Monoclonal Antibody Mediated by Fab-Fab Interaction

2020 ◽  
Vol 109 (1) ◽  
pp. 443-451 ◽  
Author(s):  
Lorenzo Gentiluomo ◽  
Dierk Roessner ◽  
Werner Streicher ◽  
Sujata Mahapatra ◽  
Pernille Harris ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Self Association

Synthesis and characterization of novel dibenz[a,c]anthracenedicarboxythioimides: the effect of thionation on self-assembly

RSC Advances ◽  
2016 ◽  
Vol 6 (82) ◽  
pp. 78784-78790 ◽  
Author(s):  
Katie. M. Psutka ◽  
Kenneth E. Maly
Keyword(s):  
Self Assembly ◽  
Liquid Crystalline ◽  
Synthesis And Characterization ◽  
Crystalline Phases ◽  
Liquid Crystalline Phases ◽  
Self Association

The effect of thionation on the formation of columnar liquid crystalline phases of dibenzanthracenedicarboximides as well as their self-association in solution is described.

scholarly journals A Dimerization Site at SCR-17/18 in Factor H Clarifies a New Mechanism for Complement Regulatory Control

2021 ◽  
Vol 11 ◽  
Author(s):  
Orla M. Dunne ◽  
Xin Gao ◽  
Ruodan Nan ◽  
Jayesh Gor ◽  
Penelope J. Adamson ◽  
...  
Keyword(s):  
Host Cell ◽  
Analytical Ultracentrifugation ◽  
Dissociation Constants ◽  
Alternative Pathway ◽  
Factor H ◽  
Regulatory Control ◽  
Size Exclusion ◽  
Complement Factor ◽  
Dimer Formation ◽  
Self Association

Complement Factor H (CFH), with 20 short complement regulator (SCR) domains, regulates the alternative pathway of complement in part through the interaction of its C-terminal SCR-19 and SCR-20 domains with host cell-bound C3b and anionic oligosaccharides. In solution, CFH forms small amounts of oligomers, with one of its self-association sites being in the SCR-16/20 domains. In order to correlate CFH function with dimer formation and the occurrence of rare disease-associated variants in SCR-16/20, we identified the dimerization site in SCR-16/20. For this, we expressed, in Pichia pastoris, the five domains in SCR-16/20 and six fragments of this with one-three domains (SCR-19/20, SCR-18/20, SCR-17/18, SCR-16/18, SCR-17 and SCR-18). Size-exclusion chromatography suggested that SCR dimer formation occurred in several fragments. Dimer formation was clarified using analytical ultracentrifugation, where quantitative c(s) size distribution analyses showed that SCR-19/20 was monomeric, SCR-18/20 was slightly dimeric, SCR-16/20, SCR-16/18 and SCR-18 showed more dimer formation, and SCR-17 and SCR-17/18 were primarily dimeric with dissociation constants of ~5 µM. The combination of these results located the SCR-16/20 dimerization site at SCR-17 and SCR-18. X-ray solution scattering experiments and molecular modelling fits confirmed the dimer site to be at SCR-17/18, this dimer being a side-by-side association of the two domains. We propose that the self-association of CFH at SCR-17/18 enables higher concentrations of CFH to be achieved when SCR-19/20 are bound to host cell surfaces in order to protect these better during inflammation. Dimer formation at SCR-17/18 clarified the association of genetic variants throughout SCR-16/20 with renal disease.

scholarly journals Purification and characterization of Robinia pseudoacacia seed lectins. A re-investigation

1986 ◽  
Vol 237 (2) ◽  
pp. 483-489 ◽  
Author(s):  
J Wantyghem ◽  
C Goulut ◽  
J P Frénoy ◽  
E Turpin ◽  
Y Goussault
Keyword(s):  
Analytical Ultracentrifugation ◽  
Human Peripheral Blood ◽  
Polyacrylamide Gel Electrophoresis ◽  
Robinia Pseudoacacia ◽  
Biological Properties ◽  
Peripheral Blood Lymphocytes ◽  
Carbohydrate Specificity ◽  
Purification And Characterization ◽  
Human Peripheral Blood Lymphocytes

Two lectins, RPA 1 and RPA 3, were purified from Robinia pseudoacacia seeds. These two lectins differ in their physicochemical and biological properties. By analytical ultracentrifugation the Mr values of RPA 1 and RPA 3 were estimated to be 59,000 and 105,000 respectively. From SDS/polyacrylamide-gel-electrophoresis data it was estimated that RPA 1 consisted of two subunits of Mr 34,000, and RPA 3 of two types of subunits (Mr 30,500 and 29,000). RPA 1 and RPA 3 were found to be glycoproteins of comparable amino acid composition. RPA 1 was the more highly glycosylated molecule (11.6% versus 4.3%). The carbohydrate-specificity of RPA 1 appears to be complex. RPA 3 was inhibited by N-acetyl-D-galactosamine and human alpha-glycoproteins. Both lectins exerted a mitogenic effect on human peripheral-blood lymphocytes. Concentrations between 0.5 and 1 microgram of RPA 3/ml gave optimal proliferative responses, whereas for RPA 1 concentrations higher than 10 micrograms/ml were needed for these responses.

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