Bovine interstitial retinol-binding protein (IRBP)—isolation and sequence analysis of cDNA clones, characterization and in vitro translation of mRNA

Liou, Gregory I.; Fong, Shao-Ling; Beattie, Wanda G.; Cook, Richard G.; Leone, Joseph W.; Landers, Robert A.; Alvarez, Richard A.; Wang, C.; Li, Y.; Bridges, C.D.B.

doi:10.1016/0042-6989(86)90052-0

Cited by 29 publications

(4 citation statements)

References 27 publications

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“…Although 2 /zg of poly(A +) RNA was loaded in each lane, the bovine band is markedly fainter than the rat doublet because high stringency conditions were used. When an identical blot was probed with B23, a bovine IRBP eDNA (43) under the same conditions, the bovine band was more intense than the rat doublet (data not shown). This indicates that the rat IRBP probe is less sensitive for bovine IRBP mRNA due to the partial homology of the IRBP mRNAs between these two species.…”

Section: Resultsmentioning

confidence: 90%

Early expression of the gene for interphotoreceptor retinol-binding protein during photoreceptor differentiation suggests a critical role for the interphotoreceptor matrix in retinal development.

Gonzalez‐Fernandez

Healy

1990

The Journal of Cell Biology

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Abstract. Interphotoreceptor retinol-binding protein (IRBP), the major protein component of the subretinal space, is in a strategic position to mediate cellular interactions between the retinal pigmented epithelium (RPE) and the neural retina. While IRBP appears to be involved in vitamin A transport during the visual cycle in the adult, the role of this protein during eye development has not been determined. As a first step to understanding the role of IRBP during retinal development, we have studied the expression of the mRNA for this glycolipoprotein during photoreceptor differentiation in the rat. A rat neural retina cDNA library was prepared from which an IRBP clone was isolated. The clone contains an open reading frame followed by a 3' noncoding sequence ending in 10 adenosine residues. The coding region has an identity of 83.9 and 82.5 % with the nucelotide sequence of human and bovine IRBP, respectively. Rats (Sprague-Dawley, Wistar, and Royal College of Surgeon pink-eyed controis) have a 6.4 and a 5.2-kb mRNA for IRBP which are present in a 1:4 ratio and thus are the only vertebrate known to definitely have more than one major form of the IRBP message. Genomic Southern blots are consistent with the hypothesis that there is only one allele of the IRBP gene, suggesting that the two forms are produced by alternative processing of the mRNA. To generate an antisense RNA probe for use in molecular titration assays and Northern blots, an Eco RI-Bam HI fragment from the coding region was subcloned in between flanking Sp6 and T7 promoters. Total RNA was prepared from undissected rat globes from postnatal days p0-p22. The expression of the mRNA for IRBP was studied by Northern blots and the level of the transcripts determined by solution hybridization assays. Approximately 1@ IRBP mRNA transcripts//~g total eye RNA are present at birth. This increases to a final level of 3.1 x 1@ transcripts/t~g total RNA by p9. The one-half maximal level of the mRNA occurs at p4.2 which is 2 wk before the onehalf maximal level of IRBP is reached in the subretinal space (Gonzalez-Fernandez, E, R. A. Landers, P. A. Glazebrook, S.-L. Fong, G. I. Liou, D. M. K. Lam, and C. D. B. . J. Cell Biol. 99:2092-2098. The expression of the mRNA for IRBP reflects the developmental emergence of the interphotoreceptor matrix as an important structure within the retina. The absolute level of IRBP is related to the volume of the subretinal space, which expands during development due to the elongation of the outer segments. The unexpected early expression of the mRNA for IRBP during retinal development suggests that the interphotoreceptor matrix is important not only to the adult during the visual cycle, but also plays a critical role during retinal development, perhaps facilitating the diffusion of hydrophobic nutrients and morphogens such as vitamin A from the RPE/choroid complex to the growing and differentiating neural retina.T H~ interphotoreceptor matrix fills the subretinal space, separating the retinal pigmented epithelium (RPE) ~ and choro...

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Section: Resultsmentioning

confidence: 90%

Early expression of the gene for interphotoreceptor retinol-binding protein during photoreceptor differentiation suggests a critical role for the interphotoreceptor matrix in retinal development.

Gonzalez‐Fernandez

Healy

1990

The Journal of Cell Biology

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“…Native IRBP was extracted from the soluble IPM fraction of bovine retinas and purified by a combination of concanavalin-A affinity, ion-exchange, and S-300 size-exclusion chromatography (53,54). The concentration of the purified IRBP was determined by absorbance spectroscopy and amino acid analysis.…”

Section: Methodsmentioning

confidence: 99%

Interphotoreceptor retinoid–binding protein removes all-trans-retinol and retinal from rod outer segments, preventing lipofuscin precursor formation

Chen

Adler

Goletz

et al. 2017

Journal of Biological Chemistry

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Interphotoreceptor retinoid-binding protein (IRBP) is a specialized lipophilic carrier that binds the all- and 11- isomers of retinal and retinol, and this facilitates their transport between photoreceptors and cells in the retina. One of these retinoids, all-retinal, is released in the rod outer segment by photoactivated rhodopsin after light excitation. Following its release, all-retinal is reduced by the retinol dehydrogenase RDH8 to all-retinol in an NADPH-dependent reaction. However, all-retinal can also react with outer segment components, sometimes forming lipofuscin precursors, which after conversion to lipofuscin accumulate in the lysosomes of the retinal pigment epithelium and display cytotoxic effects. Here, we have imaged the fluorescence of all-retinol, all-retinal, and lipofuscin precursors in real time in single isolated mouse rod photoreceptors. We found that IRBP removes all-retinol from individual rod photoreceptors in a concentration-dependent manner. The rate constant for retinol removal increased linearly with IRBP concentration with a slope of 0.012 min μm IRBP also removed all-retinal, but with much less efficacy, indicating that the reduction of retinal to retinol promotes faster clearance of the photoisomerized rhodopsin chromophore. The presence of physiological IRBP concentrations in the extracellular medium resulted in lower levels of all-retinal and retinol in rod outer segments following light exposure. It also prevented light-induced lipofuscin precursor formation, but it did not remove precursors that were already present. These findings reveal an important and previously unappreciated role of IRBP in protecting the photoreceptor cells against the cytotoxic effects of accumulated all-retinal.

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“…IRBP is a single large polypeptide of about 140 kDa Wiggert et al, 1986) and is the principal soluble protein of the IPM (Pfeffer et al, 1983). The gene ) and cDNA (Barrett et al, 1985;Liou et al, 1986;Redmond et al, 1989) for bovine IRBP have been cloned and the sequences of these clones have been determined. The deduced polypeptide sequence of bovine IRBP contains 1264 aa and five putative glycosylation sites, totaling about 147 kDa.…”

Section: Introductionmentioning

confidence: 99%

Cloning of cDNAs encoding human interphotoreceptor retinoid-binding protein (IRBP) and comparison with bovine IRBP sequences

Redmond

et al. 1989

Gene

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We have determined the sequence of the human interphotoreceptor retinoid-binding protein mRNA from three separately isolated cDNAs. The sequence is 4.28 kb long and encodes a protein of 1247 amino acids (aa) including a putative signal peptide and propeptide. The sequence is shorter (by about 1.67 kb) than the bovine mRNA with the major difference in the lengths located in the 3'-untranslated region. We suggest that this resulted from an insertion in the bovine gene or a large deletion from the human gene. The insertion/deletion is flanked on either side by sequences that are similar in the bovine and human sequences. Like the bovine polypeptide, the deduced protein sequence from the human cDNA contains a fourfold repeat, with each repeat containing about 300 aa. Among the four repeats, the identity is about 30-40%. The identity between the complete bovine and human polypeptide sequences is 84%. The identity between the nucleotide sequences is 83% (excluding the major insertion/deletion). Comparison with the bovine gene indicates that the human sequence may lack about 5-10 bp at the 5' end of the cDNA; it, however, includes a poly(A) tail at the 3' end. Thus, the human sequence is virtually full length, is similar to the bovine sequence, and contains a striking fourfold repeat.

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Bovine interstitial retinol-binding protein (IRBP)—isolation and sequence analysis of cDNA clones, characterization and in vitro translation of mRNA

Cited by 29 publications

References 27 publications

Early expression of the gene for interphotoreceptor retinol-binding protein during photoreceptor differentiation suggests a critical role for the interphotoreceptor matrix in retinal development.

Early expression of the gene for interphotoreceptor retinol-binding protein during photoreceptor differentiation suggests a critical role for the interphotoreceptor matrix in retinal development.

Interphotoreceptor retinoid–binding protein removes all-trans-retinol and retinal from rod outer segments, preventing lipofuscin precursor formation

Cloning of cDNAs encoding human interphotoreceptor retinoid-binding protein (IRBP) and comparison with bovine IRBP sequences

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