title: Steric volume exclusion sets soluble protein concentrations in photoreceptor sensory cilia.
abstract: Proteins segregate into discrete subcellular compartments via a variety of mechanisms, including motor protein transport, local binding, and diffusion barriers. This physical separation of cell functions serves, in part, as a mechanism for controlling compartment activity by allowing regulation of local protein concentrations. In this study we explored how soluble protein size impacts access to the confined space within the retinal photoreceptor outer segment signaling compartment and discovered a strikingly steep relationship. We find that GFP monomers, dimers, and trimers expressed transgenically in frog rods are present in the outer segment at 1.8-, 2.9-, and 6.8-fold lower abundances, relative to the cell body, than the small soluble fluorescent marker, calcein. Theoretical analysis, based on statistical-mechanical models of molecular access to polymer meshes, shows that these observations can be explained by the steric hindrance of molecules occupying the highly constrained spaces between outer segment disc membranes. This mechanism may answer a long-standing question of how the soluble regulatory protein, arrestin, is effectively excluded from the outer segments of dark-adapted rods and cones. Generally, our results suggest an alternate mode for the control of protein access to cell domains based on dynamic, size-dependent compartmental partitioning that does not require diffusion barriers, active transport, or large numbers of immobile binding sites.
authors.affiliation: Department of Ophthalmology, State University of New York Upstate Medical University, Syracuse, NY 13210, USA.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Najafi
authors.forename: Nycole A
authors.initials: NA
authors.lastname: Maza
authors.forename: Peter D
authors.initials: PD
authors.lastname: Calvert
mesh_terms: D000818:Animals; D002451:Cell Compartmentation; D002923:Cilia / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D008954:Models, Biological; D017948:Retinal Rod Photoreceptor Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure*; D012995:Solubility; D014982:Xenopus laevis
chemical_list: D049452:Green Fluorescent Proteins