Dimerization of human recombinant resistin involves covalent and noncovalent interactions

Area of Science:

• Biochemistry
• Endocrinology
• Molecular Biology

Background:

• Resistin is an adipocyte-secreted hormone implicated in obesity and type 2 diabetes in mice.
• Human resistin's physiological role remains unclear despite protein similarities with mouse resistin.
• Understanding human resistin structure and function is crucial for metabolic disease research.

Purpose of the Study:

• To purify and refold human recombinant resistin.
• To investigate the structural properties and dimerization of human resistin.
• To compare human resistin's characteristics with those of mouse resistin.

Main Methods:

• Purification and refolding of human recombinant resistin.
• Gel filtration and SDS-PAGE (reducing and non-reducing) to analyze protein dimerization.
• Circular dichroism spectroscopy to determine secondary structure.

Main Results:

• Two refolding methods successfully produced human recombinant resistin.
• Human resistin exhibits a tendency to form dimers, similar to mouse resistin.
• Dimerization involves both disulfide (covalent) and non-covalent interactions.
• Secondary structure analysis shows a mix of alpha-helical, beta-sheet, and unordered conformations.

Conclusions:

• Human resistin can be purified and refolded, forming dimers via covalent and non-covalent interactions.
• Cysteine 22 in human resistin may stabilize dimers through disulfide bonds.
• These findings provide insights into human resistin structure and potential function in metabolic processes.