3 research outputs found
The Membrane-binding Motif of the Chloroplast Signal Recognition Particle Receptor (cpFtsY) Regulates GTPase Activity*S⃞♦
The chloroplast signal recognition particle (cpSRP) and its receptor
(cpFtsY) function in thylakoid biogenesis to target integral membrane proteins
to thylakoids. Unlike cytosolic SRP receptors in eukaryotes, cpFtsY partitions
between thylakoid membranes and the soluble stroma. Based on sequence
alignments, a membrane-binding motif identified in Escherichia coli
FtsY appears to be conserved in cpFtsY, yet whether the proposed motif is
responsible for the membrane-binding function of cpFtsY has yet to be shown
experimentally. Our studies show that a small N-terminal region in cpFtsY
stabilizes a membrane interaction critical to cpFtsY function in
cpSRP-dependent protein targeting. This membrane-binding motif is both
necessary and sufficient to direct cpFtsY and fused passenger proteins to
thylakoids. Our results demonstrate that the cpFtsY membrane-binding motif may
be functionally replaced by the corresponding region from E. coli,
confirming that the membrane-binding motif is conserved among organellar and
prokaryotic homologs. Furthermore, the capacity of cpFtsY for lipid binding
correlates with liposome-induced GTP hydrolysis stimulation. Mutations that
debilitate the membrane-binding motif in cpFtsY result in higher rates of GTP
hydrolysis, suggesting that negative regulation is provided by the intact
membrane-binding region in the absence of a bilayer. Furthermore, NMR and CD
structural studies of the N-terminal region and the analogous region in the
E. coli SRP receptor revealed a conformational change in secondary
structure that takes place upon lipid binding. These studies suggest that the
cpFtsY membrane-binding motif plays a critical role in the intramolecular
communication that regulates cpSRP receptor functions at the membrane