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Prediktion och mutation av fatty acid binding protein, FABP

Explore the folding, stability, and binding properties of Fatty Acid Binding Protein (FABP) through site-specific mutagenesis. Study the variant characteristics of FABP and its potential applications in protein chemistry courses.

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Prediktion och mutation av fatty acid binding protein, FABP

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  1. Prediktion och mutation av fatty acid binding protein, FABP

  2. 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 Fatty acid binding protein FABP

  3. Folding of FABP FABP denature following Two-state process

  4. FABP from Saharan desert ant Cataglyphis fortis • Why study cataglyphis?? • Neither Xray structure is known or ANY stability measurement have ever been performed earlier

  5. FABP råtta FABP Cataglyphis

  6. Sekvens FABP

  7. Site-specific mutagenesis • Introduce Trp in Fatty acid binding protein • Use the constructs for characterization of the variants in courses in protein chemistry

  8. Fatty acid binding protein (FABP)

  9. QuikChange

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

  11. Characterization of FABP • CD spectra • Stability • Binding properties • Folding/refolding

  12. CataglyphisThe most thermotolerant eukaryotic species in the world

  13. Riktad mutagenes

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