Supported planar bilayers for study of the immunological synapse

Area of Science:

• Immunology
• Biophysics
• Cell Biology

Background:

• Supported planar bilayers have been instrumental in immunology for over 25 years.
• The technique has evolved to study MHC-peptide complex and adhesion molecule activity.
• Recent advancements allow for measuring 2D affinities and immunological synapse formation.

Purpose of the Study:

• To detail methods for incorporating various proteins into supported planar bilayers.
• To highlight the adaptability of existing reagents for planar bilayer presentation.
• To emphasize the physiological relevance of mobile proteins on supported bilayers.

Main Methods:

• Incorporation of glycolipid-anchored membrane proteins, 6-histidine-tagged soluble proteins, and monobiotinylated soluble proteins.
• Adaptation of reagents from MHC-peptide tetramer staining for planar bilayer use.
• Utilizing supported planar bilayers for studying laterally mobile proteins.

Main Results:

• Demonstrated successful incorporation of diverse protein types into supported planar bilayers.
• Showcased the adaptability of MHC-peptide tetramer reagents for this system.
• Established supported planar bilayers as a platform for mobile protein presentation.

Conclusions:

• Supported planar bilayers provide a physiologically relevant model for cell-surface molecule interactions.
• The lateral mobility of proteins on these bilayers aids in visualizing cellular rearrangements.
• This approach enhances the study of immune cell interactions and the immunological synapse.