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Journal article
Retinal degeneration 3 (RD3) protein, a retinal guanylyl cyclase regulator, forms a monomeric and elongated four-helix bundle
The Journal of biological chemistry, v 294(7), pp 2318-2328
15 Feb 2019
PMID: 30559291
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Retinal degeneration 3 (RD3) protein promotes accumulation of retinal membrane guanylyl cyclase (RetGC) in the photoreceptor outer segment and suppresses RetGC activation by guanylyl cyclase-activating proteins (GCAPs). Mutations truncating RD3 cause severe congenital blindness by preventing the inhibitory binding of RD3 to the cyclase. The high propensity of RD3 to aggregate in solution has prevented structural analysis. Here, we produced a highly soluble variant of human RD3 (residues 18-160) that is monomeric and can still bind and negatively regulate RetGC. The NMR solution structure of RD3 revealed an elongated backbone structure (70 A long and 30 A wide) consisting of a four-helix bundle with a long unstructured loop between helices 1 and 2. The structure reveals that RD3 residues previously implicated in the RetGC binding map to a localized and contiguous area on the structure, involving a loop between helices 2 and 3 and adjacent parts of helices 3 and 4. The NMR structure of RD3 was validated by mutagenesis. Introducing Trp(85) or Phe(29) to replace Cys or Leu, respectively, disrupts packing in the hydrophobic core and lowers RD3's apparent affinity for RetGC1. Introducing a positive charge at the interface (Glu(32) to Lys) also lowered the affinity. Conversely, introducing Val in place of Cys(93) stabilized the hydrophobic core and increased the RD3 affinity for the cyclase. The NMR structure of RD3 presented here provides a structural basis for elucidating RD3-RetGC interactions relevant for normal vision or blindness.
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Details
- Title
- Retinal degeneration 3 (RD3) protein, a retinal guanylyl cyclase regulator, forms a monomeric and elongated four-helix bundle
- Creators
- Igor V. Peshenko - Salus UniversityQinhong Yu - University of California, DavisSunghyuk Lim - University of California, DavisDiana Cudia - University of California, DavisAlexander M. Dizhoor - Salus UniversityJames B. Ames - University of California, Davis
- Publication Details
- The Journal of biological chemistry, v 294(7), pp 2318-2328
- Publisher
- Amer Soc Biochemistry Molecular Biology Inc
- Number of pages
- 11
- Grant note
- R01EY011522 / NATIONAL EYE INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Eye Institute (NEI) EY11522; EY012347 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA Pennsylvania Department of Health CURE Formula grant
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Neurobiology and Anatomy; Pennsylvania College of Optometry (PCO)
- Web of Science ID
- WOS:000458895400010
- Scopus ID
- 2-s2.0-85061575985
- Other Identifier
- 991022035111704721
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- Web of Science research areas
- Biochemistry & Molecular Biology