Comparison of the molecular properties of retinitis pigmentosa P23H and N15S amino acid replacements in rhodopsin.

Mitchell, James

Balem, Fernanda

Tirupula, Kalyan

Man, David

Dhiman, Harpreet Kaur

Yanamala, Naveena

Ollesch, Julian

Planas-Iglesias, Joan

Jennings, Barbara J

Gerwert, Klaus

Iannaccone, Alessandro

Klein-Seetharaman, Judith

2019-07-01T13:58:09Z

2019-07-01T13:58:09Z

2019-01

2019-07-01T13:58:07Z

Mutations in the RHO gene encoding for the visual pigment protein, rhodopsin, are among the most common cause of autosomal dominant retinitis pigmentosa (ADRP). Previous studies of ADRP mutations in different domains of rhodopsin have indicated that changes that lead to more instability in rhodopsin structure are responsible for more severe disease in patients. Here, we further test this hypothesis by comparing side-by-side and therefore quantitatively two RHO mutations, N15S and P23H, both located in the N-terminal intradiscal domain. The in vitro biochemical properties of these two rhodopsin proteins, expressed in stably transfected tetracycline-inducible HEK293S cells, their UV-visible absorption, their Fourier transform infrared, circular dichroism and Metarhodopsin II fluorescence spectroscopy properties were characterized. As compared to the severely impaired P23H molecular function, N15S is only slightly defective in structure and stability. We propose that the molecular basis for these structural differences lies in the greater distance of the N15 residue as compared to P23 with respect to the predicted rhodopsin folding core. As described previously for WT rhodopsin, addition of the cytoplasmic allosteric modulator chlorin e6 stabilizes especially the P23H protein, suggesting that chlorin e6 may be generally beneficial in the rescue of those ADRP rhodopsin proteins whose stability is affected by amino acid replacement.

PONE-D-17-44536

1932-6203

1932-6203

https://hdl.handle.net/10161/19055

eng

Public Library of Science (PLoS)

PloS one

10.1371/journal.pone.0214639

Science & Technology

Multidisciplinary Sciences

Science & Technology - Other Topics

POINT MUTATION

VITAMIN-A

MUTANTS

GENE

EXPRESSION

SUPPLEMENTATION

GLYCOSYLATION

IMPROVEMENT

STABILITY

BINDING

Comparison of the molecular properties of retinitis pigmentosa P23H and N15S amino acid replacements in rhodopsin.

Journal article

Iannaccone, Alessandro|0000-0001-5737-8424

e0214639

5

School of Medicine

Duke

Ophthalmology, Vitreoretinal Diseases & Surgery

Ophthalmology

Clinical Science Departments

Published

14