Structural and functional characterization of recombinant human cellular retinaldehyde-binding protein

Protein Science : a Publication of the Protein Society
John W CrabbD Bok

Abstract

Cellular retinaldehyde-binding protein (CRALBP) is abundant in the retinal pigment epithelium (RPE) and Müller cells of the retina where it is thought to function in retinoid metabolism and visual pigment regeneration. The protein carries 11-cis-retinal and/or 11-cis-retinol as endogenous ligands in the RPE and retina and mutations in human CRALBP that destroy retinoid binding functionality have been linked to autosomal recessive retinitis pigmentosa. CRALBP is also present in brain without endogenous retinoids, suggesting other ligands and physiological roles exist for the protein. Human recombinant cellular retinaldehyde-binding protein (rCRALBP) has been over expressed as non-fusion and fusion proteins in Escherichia coli from pET3a and pET19b vectors, respectively. The recombinant proteins typically constitute 15-20% of the soluble bacterial lysate protein and after purification, yield about 3-8 mg per liter of bacterial culture. Liquid chromatography electrospray mass spectrometry, amino acid analysis, and Edman degradation were used to demonstrate that rCRALBP exhibits the correct primary structure and mass. Circular dichroism, retinoid HPLC, UV-visible absorption spectroscopy, and solution state 19F-NMR were used to char...Continue Reading

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Citations

Mar 23, 2002·Protein Science : a Publication of the Protein Society·John W CrabbHenry F Hoff
Mar 5, 2008·Clinical & Developmental Immunology·Cornelia A DeegManfred Stangassinger
Apr 25, 2000·Biochemical and Biophysical Research Communications·L A LuckC H Peck
May 9, 2012·Ophthalmology·Karin W LittinkL Ingeborgh van den Born
Nov 21, 2015·Proteomics. Clinical Applications·Hannah E BowreyZsolt Ablonczy
Nov 26, 2008·Veterinary Immunology and Immunopathology·Cornelia A Deeg
Jan 22, 2003·The Journal of Biological Chemistry·Zhiping WuJohn W Crabb
Jan 22, 2003·The Journal of Biological Chemistry·Irina GolovlevaJohn W Crabb
Feb 18, 2014·British Journal of Pharmacology·Stephen P H AlexanderM Zimmermann
May 3, 2002·The Journal of Biological Chemistry·Saumendra N SarkarGanes C Sen
Nov 17, 2001·Chemical Reviews·R R Rando
Jan 11, 2018·Biochemistry·Timothy S OwenKrzysztof Palczewski

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