Denaturation of pH-dependent CRY-DASH Protein

Presentation Type

Poster

Faculty Advisor

Yvonne Gindt

Access Type

Event

Start Date

26-4-2024 12:45 PM

End Date

26-4-2024 1:44 PM

Description

Vibrio cholerae Cryptochrome-1 (VcCRY-1) is a CRY-DASH subfamily of Cryptochrome/Photolyase family of (CPF) of blue-light photoreceptor proteins. The CPF includes DNA Photolyase, which repairs UV-damaged DNA through blue light, Cryptochrome, which signals for growth and development through blue light, and CRY-DASH, which carries out both roles in vivo. Under high light intensity, CRY-DASH will tend to behave as DNA photolyase and repair UV-damaged DNA using blue light. Under low light intensity, it will tend to behave as Cryptochromes and serve as blue light signaling proteins. It appears that substrate binding and pH changes prevent the formation of the FAD state with the loss of the proton acceptor. We will test for temperature and pH-sensitive structural changes using the emission of the FAD cofactor. Experiments were designed to use the emission of the FAD cofactor to report oligomeric changes as a function of temperature. Results show that the protein at a lower pH requires a higher temperature for thermal denaturation. Therefore, protein structure appears to be significantly more stable at lower pH.

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Apr 26th, 12:45 PM Apr 26th, 1:44 PM

Denaturation of pH-dependent CRY-DASH Protein

Vibrio cholerae Cryptochrome-1 (VcCRY-1) is a CRY-DASH subfamily of Cryptochrome/Photolyase family of (CPF) of blue-light photoreceptor proteins. The CPF includes DNA Photolyase, which repairs UV-damaged DNA through blue light, Cryptochrome, which signals for growth and development through blue light, and CRY-DASH, which carries out both roles in vivo. Under high light intensity, CRY-DASH will tend to behave as DNA photolyase and repair UV-damaged DNA using blue light. Under low light intensity, it will tend to behave as Cryptochromes and serve as blue light signaling proteins. It appears that substrate binding and pH changes prevent the formation of the FAD state with the loss of the proton acceptor. We will test for temperature and pH-sensitive structural changes using the emission of the FAD cofactor. Experiments were designed to use the emission of the FAD cofactor to report oligomeric changes as a function of temperature. Results show that the protein at a lower pH requires a higher temperature for thermal denaturation. Therefore, protein structure appears to be significantly more stable at lower pH.