Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Method Development for Determining the Stability of Heat Stable Proteins Combined with Biophysical Characterization of Human Calmodulin and the Disease Associated Variant D130G
Linköping University, Department of Physics, Chemistry and Biology.
Linköping University, Department of Physics, Chemistry and Biology.
Linköping University, Department of Physics, Chemistry and Biology.
Linköping University, Department of Physics, Chemistry and Biology.
Show others and affiliations
2016 (English)Independent thesis Basic level (degree of Bachelor), 10,5 credits / 16 HE creditsStudent thesis
Abstract [en]

Calmodulin is a highly conserved calcium ion binding protein expressed in all eukaryotic species. The 149 amino acid residues in the primary structure are organized in seven α helices with the highly flexible central α helix connecting the two non-cooperative domains of calmodulin. Each domain contains two EF-hand motifs to which calcium ions bind in a cooperative manner, hence the binding of four calcium ions saturate one calmodulin molecule. In the cardiovascular area calmodulin is involved in the activation of cardiac muscle contraction, and mutations that arise in the genetic sequence of the protein often have severe consequences. One such consequential mutation that can arise brings about the replacement of the highly conserved aspartic acid with glycine at position 130 in the amino acid sequence. In this research, the thermal and chemical stability within the C domain of the D130G variant of human calmodulin was investigated using a new method only requiring circular dichroism spectroscopic measurements. Affinity studies within the C domain of the D130G variant of human calmodulin were performed using fluorescence spectroscopy, and the ligands chosen for this purpose were trifluoperazine and p- HTMI. All analytical experiments were performed with the C domain of wild type human calmodulin as a reference. From the new method, it was concluded that the C domain of the D130G variant of human calmodulin has a slightly decreased stability in terms of Tm and Cm values compared to the C domain of wild type human calmodulin. The affinity analyses indicated that neither trifluoperazine nor p-HTMI discriminates between the C domain of the D130G variant of human calmodulin and the C domain of wild type human calmodulin in terms of dissociation constants. The pivotal outcome from this research is that the new method is applicable for determination of the stability parameters Tm and Cm of heat stable proteins. 

Place, publisher, year, edition, pages
2016. , 47 p.
Keyword [en]
Calmodulin, wild type, D130G variant, method, affinity, trifluoperazine, p-HTMI.
National Category
Physical Chemistry Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:liu:diva-138022ISRN: LITH-IFM-G-EX--16/3178--SEOAI: oai:DiVA.org:liu-138022DiVA: diva2:1106033
Subject / course
Chemical Biology
Presentation
2016-05-27, ACAS, Linköpings Universitet (campus valla), A-huset, Linköping, 08:15 (Swedish)
Supervisors
Examiners
Available from: 2017-06-09 Created: 2017-06-06 Last updated: 2017-06-09Bibliographically approved

Open Access in DiVA

fulltext(3606 kB)20 downloads
File information
File name FULLTEXT01.pdfFile size 3606 kBChecksum SHA-512
cfa4d603fce78d2bc48eed6a32f907d20ac695d0bd8e046b63a20576322a543cf213f89f15ac107bf1d9d7f05569f0ea81ad6f72314c8c7a2db71cc166772c3d
Type fulltextMimetype application/pdf

Other links

http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-138022

Search in DiVA

By author/editor
Aleckovic, EhlimanaChamoun, SherleyEinarsson, Ellen
By organisation
Department of Physics, Chemistry and Biology
Physical ChemistryBiochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 20 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 98 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf