Equilibrium Refolding Transitions Driven by Trifluoroethanol and by Guanidine Hydrochloride Dilution Are Similar in GTPase Effector Domain: Implications to Sequence−Self-Association Paradigm†

Biochemistry ◽  
2008 ◽  
Vol 47 (49) ◽  
pp. 12945-12953 ◽  
Author(s):  
Jeetender Chugh ◽  
Shilpy Sharma ◽  
Ramakrishna V. Hosur
Keyword(s):  
Guanidine Hydrochloride ◽  
Effector Domain ◽  
Gtpase Effector Domain ◽  
Self Association

scholarly journals NMR Derived Model of GTPase Effector Domain (GED) Self Association: Relevance to Dynamin Assembly

PLoS ONE ◽  
2012 ◽  
Vol 7 (1) ◽  
pp. e30109
Author(s):  
Swagata Chakraborty ◽  
Supriya Pratihar ◽  
Ramakrishna V. Hosur
Keyword(s):  
Effector Domain ◽  
Gtpase Effector Domain ◽  
Self Association

scholarly journals Macromolecular properties and polymeric structure of canine tracheal mucins

1991 ◽  
Vol 276 (2) ◽  
pp. 525-532 ◽  
Author(s):  
V Shankar ◽  
A K Virmani ◽  
B Naziruddin ◽  
G P Sachdev
Keyword(s):  
Gel Electrophoresis ◽  
Guanidine Hydrochloride ◽  
Polyacrylamide Gel Electrophoresis ◽  
Hydrophobic Properties ◽  
Experimental Conditions ◽  
Protein Core ◽  
Polymeric Structure ◽  
Composite Gel ◽  
Self Association ◽  
Static Laser Light Scattering

Two high-Mr mucus glycoproteins (mucins), CTM-A and CTM-B, were highly purified from canine tracheal pouch secretions, and their macromolecular properties as well as polymeric structure were investigated. On SDS/composite-gel electrophoresis, a diffuse band was observed for each mucin. Polyacrylamide-gel electrophoresis using 6% gels also showed the absence of low-Mr contaminants in the mucins. Comparison of chemical and amino acid compositions revealed significant differences between the two mucins. Using a static-laser-light-scattering technique, CTM-A and CTM-B were found to have weight-average Mr values of about 11.0 x 10(6) and 1.4 x 10(6) respectively. Both mucins showed concentration-dependent aggregation in buffer containing 6 M-guanidine hydrochloride. Under similar experimental conditions, reduced-alkylated CTM-A had an Mr of 5.48 x 10(6) and showed no concentration-dependent aggregation. Hydrophobic properties of the mucins, investigated by the fluorescent probe technique using mansylphenylalanine as the probe, showed the presence of a large number of low-affinity (KD approx. 10(5) M) binding sites. These sites appeared to be located on the non-glycosylated regions of the protein core, since Pronase digestion of the mucins almost completely eliminated probe binding. Reduction of disulphide bonds of CTM-A and CTM-B did not significantly alter the probe-binding properties. Also, addition of increasing NaCl concentrations (0.03-1.0 M) to the buffer caused only a small change in the hydrophobic properties of native and reduced-alkylated mucins. CTM-A was deglycosylated, without notable in the hydrophobic properties of native and reduced-alkylated mucins. CTM-A was deglycosylated, without notable degradation, using a combination of chemical and enzymic methods. On SDS/PAGE the protein core was estimated to have an Mr of approx. 60,000. On the basis of the protein and carbohydrate contents of the major mucin CTM-A, the mucin monomer was calculated to have an Mr of approx. 140,000. The high Mr (11 x 10(6] observed by physical methods is therefore due to self-association of the mucin monomer subunits.

scholarly journals De novo missense variant in the GTPase effector domain (GED) of DNM1L leads to static encephalopathy and seizures

2019 ◽  
Vol 5 (3) ◽  
pp. a003673 ◽  
Author(s):  
Nurit Assia Batzir ◽  
Pranjali K. Bhagwat ◽  
Tanya N. Eble ◽  
Pengfei Liu ◽  
Christine M. Eng ◽  
...  
Keyword(s):  
De Novo ◽  
Missense Variant ◽  
Effector Domain ◽  
Gtpase Effector Domain ◽  
Static Encephalopathy

1H, 15N, 13C resonance assignment of 9.7 M urea-denatured state of the GTPase effector domain (GED) of dynamin

2008 ◽  
Vol 3 (1) ◽  
pp. 13-16 ◽  
Author(s):  
Jeetender Chugh ◽  
Shilpy Sharma ◽  
Dinesh Kumar ◽  
Ramakrishna V. Hosur
Keyword(s):  
Resonance Assignment ◽  
Effector Domain ◽  
Denatured State ◽  
Gtpase Effector Domain

scholarly journals Adult-onset autosomal dominant spastic paraplegia linked to a GTPase-effector domain mutation of dynamin 2

BMC Neurology ◽  
2015 ◽  
Vol 15 (1) ◽  
Author(s):  
Nyamkhishig Sambuughin ◽  
Lev G. Goldfarb ◽  
Tatiana M. Sivtseva ◽  
Tatiana K. Davydova ◽  
Vsevolod A. Vladimirtsev ◽  
...  
Keyword(s):  
Autosomal Dominant ◽  
Adult Onset ◽  
Spastic Paraplegia ◽  
Effector Domain ◽  
Gtpase Effector Domain ◽  
Dynamin 2

scholarly journals Intrapolypeptide Interactions between the GTPase Effector Domain (GED) and the GTPase Domain Form the Bundle Signaling Element in Dynamin Dimers

Biochemistry ◽  
2014 ◽  
Vol 53 (36) ◽  
pp. 5724-5726 ◽  
Author(s):  
Saipraveen Srinivasan ◽  
Juha-Pekka Mattila ◽  
Sandra L. Schmid
Keyword(s):  
Gtpase Domain ◽  
Effector Domain ◽  
Gtpase Effector Domain

scholarly journals The GTPase Effector Domain Sequence of the Dnm1p GTPase Regulates Self-Assembly and Controls a Rate-limiting Step in Mitochondrial Fission

2001 ◽  
Vol 12 (9) ◽  
pp. 2756-2766 ◽  
Author(s):  
Noelle H. Fukushima ◽  
Ellen Brisch ◽  
Brian R. Keegan ◽  
William Bleazard ◽  
Janet M. Shaw
Keyword(s):  
Mitochondrial Fission ◽  
Yeast Two Hybrid ◽  
Effector Domain ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Gtpase Effector Domain ◽  
Rate Limiting ◽  
Two Hybrid

Dnm1p belongs to a family of dynamin-related GTPases required to remodel different cellular membranes. In budding yeast, Dnm1p-containing complexes assemble on the cytoplasmic surface of the outer mitochondrial membrane at sites where mitochondrial tubules divide. Our previous genetic studies suggested that Dnm1p's GTPase activity was required for mitochondrial fission and that Dnm1p interacted with itself. In this study, we show that bacterially expressed Dnm1p can bind and hydrolyze GTP in vitro. Coimmunoprecipitation studies and yeast two-hybrid analysis suggest that Dnm1p oligomerizes in vivo. With the use of the yeast two-hybrid system, we show that this Dnm1p oligomerization is mediated, in part, by a C-terminal sequence related to the GTPase effector domain (GED) in dynamin. The Dnm1p interactions characterized here are similar to those reported for dynamin and dynamin-related proteins that form higher order structures in vivo, suggesting that Dnm1p assembles to form rings or collars that surround mitochondrial tubules. Based on previous findings, a K705A mutation in the Dnm1p GED is predicted to interfere with GTP hydrolysis, stabilize active Dnm1p-GTP, and stimulate a rate-limiting step in fission. Here we show that expression of the Dnm1 K705A protein in yeast enhances mitochondrial fission. Our results provide evidence that the GED region of a dynamin-related protein modulates a rate-limiting step in membrane fission.

scholarly journals Self-association of proteoglycan subunits from pig laryngeal cartilage

1978 ◽  
Vol 171 (1) ◽  
pp. 109-114 ◽  
Author(s):  
J K Sheehan ◽  
I A Nieduszynski ◽  
C F Phelps
Keyword(s):  
Light Scattering ◽  
Ionic Strength ◽  
Guanidine Hydrochloride ◽  
Sedimentation Velocity ◽  
Dilute Solution ◽  
Thiol Groups ◽  
Dimer Model ◽  
Laryngeal Cartilage ◽  
Self Association ◽  
Corroborative Evidence

Proteoglycans from pig laryngeal cartilage prepared by dissociative extraction in guanidine hydrochloride were studied in dilute solution by light-scattering and ultracentrifugation. In buffered 150mM-NaCl, pH7.4, the proteoglycan particle weights were about 5×10(6) daltons, but at 100mM-, 200mM- and 300mM-NaCl particle weights of 2.5×10(6)–3.0×10(6) daltons were observed. These results, together with corroborative evidence from sedimentation-velocity experiments, were interpreted in terms of proteoglycans self-associating at physiological ionic strength. The data were examined by using a proteoglycan monomer-dimer model. Proteoglycan preparations that had thiol groups partially carboxymethylated gave particle weights of 3.2×10(6)–3.5×10(6) daltons in 150mM-NaCl, which suggested that carboxymethylation inhibited multimerization and hence that the protein core is implicated in the binding site. Further studies showed that the multimers were stable to 60 degrees C, unlike the hyaluronate-proteoglycan complex.

The self-association of glycosyl phenylazo dyes (Yariv antigens)

1978 ◽  
Vol 31 (10) ◽  
pp. 2225 ◽  
Author(s):  
EF Woods ◽  
GG Lilley ◽  
MA Jermyn
Keyword(s):  
Guanidine Hydrochloride ◽  
Sodium Dodecyl ◽  
Entropy Change ◽  
Wide Distribution ◽  
Enthalpy Change ◽  
Spectral Measurements ◽  
Concentrated Solutions ◽  
Weight Concentration ◽  
Equilibrium Sedimentation ◽  
Self Association

The aggregation of tris(4-glycosyloxyphenylazo)phloroglucinol compounds (Yariv antigens) where glyco = β-D-gluco, β-D-manno, β-D-galacto and α- D-galacto has been investigated by equilibrium sedimentation, the photoelectric scanning absorption optical system being utilized. The compounds have been shown to undergo a strong self-association in water and an analysis procedure employing Laplace transforms indicated a very wide distribution of species in the ultracentrifuge cell at equilibrium. To determine the type of association the concentration of monomer as a function of total weight concentration was calculated by using the recently developed Ω function. When the concentration of monomer is very low, as in the present case, the analysis is difficult because very slight curvature in the Ω-function plot has a large effect on the extrapolation to zero concentration. ��� The results are best described by an isodesmic indefinite self- association. The β-glucoside Yariv antigen was the most extensively investigated and the most highly purified sample was shown to have an association constant of 2.5 x 107 l. mol-1. The order of the association constants of the Yariv compounds was found to be β-glucoside > β- galactoside > α-galactoside > β-mannoside. For the β-glucoside compound an increase in temperature favours species of lower molecular weight, and from the enthalpy change a positive entropy change for the association is indicated. The compounds are disaggregated by concentrated solutions of urea and guanidine hydrochloride. Spectral measurements on the β-glucoside compound indicate disaggregation by sodium dodecyl sulfate above the critical micelle concentration.

Sign in / Sign up

Export Citation Format

Share Document