Engineering visual arrestin-1 with special functional characteristics.

Vishnivetskiy SA, Chen Q, Palazzo MC, Brooks EK, Altenbach C, Iverson TM, Hubbell WL, Gurevich VV
J Biol Chem. 2013 288 (5): 3394-405

PMID: 23250748 · PMCID: PMC3561558 · DOI:10.1074/jbc.M112.445437

Arrestin-1 preferentially binds active phosphorylated rhodopsin. Previously, a mutant with enhanced binding to unphosphorylated active rhodopsin (Rh*) was shown to partially compensate for lack of rhodopsin phosphorylation in vivo. Here we showed that reengineering of the receptor binding surface of arrestin-1 further improves the binding to Rh* while preserving protein stability. In mammals, arrestin-1 readily self-associates at physiological concentrations. The biological role of this phenomenon can only be elucidated by replacing wild type arrestin-1 in living animals with a non-oligomerizing mutant retaining all other functions. We demonstrate that constitutively monomeric forms of arrestin-1 are sufficiently stable for in vivo expression. We also tested the idea that individual functions of arrestin-1 can be independently manipulated to generate mutants with the desired combinations of functional characteristics. Here we showed that this approach is feasible; stable forms of arrestin-1 with high Rh* binding can be generated with or without the ability to self-associate. These novel molecular tools open the possibility of testing of the biological role of arrestin-1 self-association and pave the way to elucidation of full potential of compensational approach to gene therapy of gain-of-function receptor mutations.

MeSH Terms (18)

Animals Arrestins beta-Arrestins Eye HEK293 Cells Humans Mice Models, Molecular Mutant Proteins Mutation Phosphates Protein Binding Protein Engineering Protein Stability Protein Structure, Tertiary Rhodopsin Static Electricity Temperature

Connections (1)

This publication is referenced by other Labnodes entities:

Links