2008
DOI: 10.1007/978-1-60327-058-8_19
|View full text |Cite
|
Sign up to set email alerts
|

Methods for Protein Characterization by Mass Spectrometry, Thermal Shift (ThermoFluor) Assay, and Multiangle or Static Light Scattering

Abstract: Mass spectrometry (MS) is widely used within structural and functional proteomics for a variety of tasks including protein quality assessment, identification, and characterization. MS is used routinely for the determination of the total mass of proteins, including N-glycosylated proteins, analysis of selenomethionine incorporation, crystal content verification, and analysis of N-glycosylation site occupancy. Protocols for sample preparation, data collection, and analysis are given.A recent development is the f… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
2
1
1
1

Citation Types

0
116
0

Year Published

2010
2010
2017
2017

Publication Types

Select...
9
1

Relationship

2
8

Authors

Journals

citations
Cited by 127 publications
(116 citation statements)
references
References 12 publications
0
116
0
Order By: Relevance
“…3A), characteristic of a protein containing 65% α-helix, 17% turns, and 8% unfolded, as calculated with DichroWeb (11) using two different algorithms, SELCON3 (12) and CONTIN (13). A ThermoFluor assay (14) was used to monitor thermal unfolding of αSyn, and to detect whether a hydrophobic core is present in the oligomer, as determined by an increase in fluorescence emitted by the dye present. We observed a sigmoidal unfolding curve for the αSyn construct, indicating a cooperative unfolding with exposure of hydrophobic residues (SI Appendix, Fig.…”
Section: Resultsmentioning
confidence: 99%
“…3A), characteristic of a protein containing 65% α-helix, 17% turns, and 8% unfolded, as calculated with DichroWeb (11) using two different algorithms, SELCON3 (12) and CONTIN (13). A ThermoFluor assay (14) was used to monitor thermal unfolding of αSyn, and to detect whether a hydrophobic core is present in the oligomer, as determined by an increase in fluorescence emitted by the dye present. We observed a sigmoidal unfolding curve for the αSyn construct, indicating a cooperative unfolding with exposure of hydrophobic residues (SI Appendix, Fig.…”
Section: Resultsmentioning
confidence: 99%
“…The thermal stabilities of wild-type hSR and the C217S, K221E, and S84G/P111L mutants were assessed using ThermoFluor, a biophysical technique for determination of relative protein stabilities based on fluorophore partition into the denatured protein upon temperature increase [28][29][30] . In the absence of SR activators, wild-type, C217S, and K221E hSR exhibited melting temperatures of 63.2 ± 0.53, 63.1 ± 0.17, and 62.7 ± 0.32°C, respectively.…”
Section: Thermal Stability Of C217s K221e and S84g/p111l Hsrmentioning
confidence: 99%
“…Binding of a Phosphate Group in Solution-We carried out DSF and CD assays (54) to investigate the binding of a phosphate in solution to the KinB-SD. The protein unfolding temperature (T m ) was measured using DSF at different phosphate concentrations.…”
Section: Resultsmentioning
confidence: 99%