Prediktion och mutation av fatty acid binding protein, FABP

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Prediktion och mutation av fatty acid binding
protein, FABP
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Fatty acid binding protein FABP

• Small 130 aa
• B-sheet protein
• Carrier protein for fatty acid and lipophilic
  substances
• Involved in transfer of fatty acid between inter
  and extracellular membranes
• Contains a hydrophobic active site with low
  specificity
• Model protein for folding studies

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Folding of FABP
FABP denature following Two-state process
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FABP from Saharan desert ant Cataglyphis fortis

• Why study cataglyphis??
• Neither Xray structure is known or ANY stability
  measurement have ever been performed earlier

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FABP råtta
FABP Cataglyphis
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Sekvens FABP
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Site-specific mutagenesis

• Introduce Trp in Fatty acid binding protein
• Use the constructs for characterization of the
  variants in courses in protein chemistry

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Fatty acid binding protein (FABP)
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QuikChange
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Purification of FABP

• Expression of FABP by IPTG
• Sonication to lyse the cells
• Purification using Qiagen NTA column

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Characterization of FABP

• CD spectra
• Stability
• Binding properties
• Folding/refolding

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Cataglyphis The most thermotolerant eukaryotic
species in the world
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Riktad mutagenes