Membrane-Bound Protein Scaffolding in Diverse Hosts Using Thylakoid Protein CURT1A

Protein scaffolding is a useful strategy
for controlling the spatial
arrangement of cellular components via protein–protein
interactions. Protein scaffolding has primarily been used to colocalize
soluble proteins in the cytoplasm, but many proteins require membrane
association for proper function. Scaffolding at select membrane domains
would provide an additional level of control over the distribution
of proteins within a cell and could aid in exploiting numerous metabolic
pathways that contain membrane-associated enzymes. We developed and
characterized a membrane-bound protein scaffolding module based on
the thylakoid protein CURT1A. This scaffolding module forms homo-oligomers
in the membrane, causing proteins fused to CURT1A to cluster together
at membrane surfaces. It is functional in diverse expression hosts
and can scaffold proteins at thylakoid membranes in chloroplasts,
endoplasmic reticulum in higher plants and Saccharomyces cerevisiae, and the inner membrane of Escherichia coli.