Sequence and expression analysis of bovine pigment epithelium-derived factor

Pérez-Mediavilla, Alberto; Chew, Christina E.; Campochiaro, Peter A.; Nickells, Robert W.; Notario, Vicente; Zack, Donald J.; Becerra, S. Patricia

doi:10.1016/s0167-4781(98)00055-4

Cited by 28 publications

(15 citation statements)

References 24 publications

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“…The size of the purified protein was determined by SDS-PAGE using several gels of a variety of polyacrylamide concentrations and under reducing conditions. The calculated size was 50 700^1000-Mr and is in agreement with the size for the native and recombinant bovine and human PEDF proteins (Wu et al, 1995, andStratikos et al, 1996;Perez-Mediavilla et al, 1998). The monkey PEDF behaved as a monomeric protein by size-exclusion ultrafiltration.…”

Section: Characterization Of Monkey Pedf Proteinsupporting

confidence: 76%

“…Biochemical and immunochemical studies have identified PEDF as a soluble extracellular protein localized in washes of the interphotoreceptor matrix (IPM), vitreous and aqueous of several mammalian species (Tombran-Tink et al, 1995;Wu et al, 1995;Ortego et al, 1996;Alberdi et al, 1998;Karakousis et al, 2001). PEDF, discovered as a product secreted by cultured RPE cells from fetal human eyes (Tombran Tink, 1991), can be released from a variety of cells, including stably transfected eukaryotic cells with full length PEDF cDNA under the control of recombinant transcriptional promoters (Stratikos et al, 1996;Perez-Mediavilla et al, 1998). By sequence homology, PEDF is a member of the serpin superfamily , formed by proteins with a common overall protein conformation with a variety of biological functions, most of which are extracellular serine protease inhibitors.…”

Section: Introductionmentioning

confidence: 99%

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Pigment epithelium-derived factor in the monkey retinal pigment epithelium and interphotoreceptor matrix: apical secretion and distribution

Becerra

Fariss

et al. 2004

Experimental Eye Research

114

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Pigment epithelium-derived factor (PEDF) is an extracellular protein derived from the retinal pigment epithelium (RPE), a tissue formed by polarized cells that release growth and trophic factors in a directional fashion. We have investigated the distribution and directional release of PEDF protein by the monkey RPE. We established primary cultures of monkey RPE cells that expressed the PEDF gene, and that synthesized and secreted the PEDF protein. Northern analysis of RPE cultures and monkey ocular tissues showed that PEDF transcripts were highly expressed in RPE as compared with several other monkey ocular tissues, being even more abundant in cultured cells than they were in the native RPE. The differentiated RPE cells in culture secreted protein that shared the immunological, biochemical and biological characteristics of PEDF. The overall PEDF levels in the RPE conditioned media reached 6·5 mg ml 2 after 8 days in culture (i.e. 1·1 pg of PEDF per RPE cell). RPE cells were cultivated on permeable supports as monolayers forming a barrier between apical and basal compartments. Apical and basal culture media were sampled at three or four-day intervals for 18 cycles, and the PEDF content was quantified. Most of the PEDF protein was significantly higher in the apical than in the basal medium (. 4 times) at the initial recovery intervals, to be detected only in the apical medium at the latter intervals. In the native monkey eye, the concentration of soluble PEDF in the interphotoreceptor matrix (144 nM) was 7-fold and 25-fold greater than in vitreous and aqueous, respectively. PEDF was abundant in the interphotoreceptor matrix surrounding rod and cone outer segments, and was detectable at lower levels in the RPE as visualized by confocal microscopy. We concluded that PEDF synthesized by the RPE is secreted preferentially from the apical surface and is distributed apically to the RPE bordering the outer segments of photoreceptors. PEDF can be a useful marker for RPE polarization and differentiation. The polarization of RPE may be an important mechanism to control PEDF secretion and our results offer interesting possibilities on regulation of PEDF. q

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Section: Characterization Of Monkey Pedf Proteinsupporting

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Pigment epithelium-derived factor in the monkey retinal pigment epithelium and interphotoreceptor matrix: apical secretion and distribution

Becerra

Fariss

et al. 2004

Experimental Eye Research

114

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“…It was discovered as a 50-kDa protein released by cultured pigment epithelial cells from fetal human retina (7). In the retina, PEDF is highly expressed in the retinal pigment epithelium (RPE) (8,9), is enriched in the interphotoreceptor matrix (10 -12), and has been implicated in photoreceptor morphogenesis (13) and in vascular and neural retinal disorders (1). It acts on photoreceptor survival (14) and in preventing the pathological invasion of neovessels (15).…”

mentioning

confidence: 99%

Identification of a Lipase-linked Cell Membrane Receptor for Pigment Epithelium-derived Factor

Notari

Baladrón

Aroca-Aguilar

et al. 2006

Journal of Biological Chemistry

Self Cite

258

295

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Pigment epithelium-derived factor (PEDF) is an extracellular multifunctional protein belonging to the serpin superfamily with demonstrable neurotrophic, gliastatic, neuronotrophic, antiangiogenic, and antitumorigenic properties. We have previously provided biochemical evidence for high affinity PEDFbinding sites and proteins in plasma membranes of retina, retinoblastoma, and CNS cells. This study was designed to reveal a receptor involved in the biological activities of PEDF. Using a yeast two-hybrid screening, we identified a novel gene from pigment epithelium of the human retina that codes for a PEDFbinding partner, which we term PEDF-R. The derived polypeptide has putative transmembrane, intracellular and extracellular regions, and a phospholipase domain. Recently, PEDF-R (TTS-2.2/independent phospholipase A 2 (PLA 2 ) and mouse desnutrin/ATGL) has been described in adipose cells as a member of the new calcium-independent PLA 2 /nutrin/patatin-like phospholipase domain-containing 2 (PNPLA2) family that possesses triglyceride lipase and acylglycerol transacylase activities. Here we describe the PEDF-R gene expression in the retina and its heterologous expression by bacterial and eukaryotic systems, and we demonstrate that its protein product has specific and high binding affinity for PEDF, has a potent phospholipase A 2 activity that liberates fatty acids, and is associated with eukaryotic cell membranes. Most importantly, PEDF binding stimulates the enzymatic phospholipase A 2 activity of PEDF-R. In conclusion, we have identified a novel PEDF-R gene in the retina for a phospholipase-linked membrane protein with high affinity for PEDF, suggesting a molecular pathway by which ligand/receptor interaction on the cell surface could generate a cellular signal.

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“…The template was plasmid phbPEDF (36), which contains the first 43 codons of the human PEDF cDNA (codons 1-20 correspond to a secretion signal peptide) followed by the 42-416 codons of the bovine PEDF cDNA. Amino acid numbers for this set of mutants refer to alignment with the human sequence.…”

Section: Methodsmentioning

confidence: 99%

Pigment Epithelium-derived Factor Binds to Hyaluronan

Becerra

Pérez-Mediavilla

Weldon

et al. 2008

Journal of Biological Chemistry

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Pigment epithelium-derived factor (PEDF) is a multifunctional serpin with antitumorigenic, antimetastatic, and differentiating activities. PEDF is found within tissues rich in the glycosaminoglycan hyaluronan (HA), and its amino acid sequence contains putative HA-binding motifs. We show that PEDF coprecipitation with glycosaminoglycans in media conditioned by human retinoblastoma Y-79 cells decreased after pretreatments with hyaluronidase, implying an association between HA and PEDF. Direct binding of human recombinant PEDF to highly purified HA was demonstrated by coprecipitation in the presence of cetylpyridinium chloride. Binding of PEDF to HA was concentration-dependent and saturable. The PEDF-HA interactions were sensitive to increasing NaCl concentrations, indicating an ionic nature of these interactions and having affinity higher than PEDF-heparin. Competition assays showed that PEDF can bind heparin and HA simultaneously. PEDF chemically modified with fluorescein retained the capacity for interacting with HA but lacked heparin affinity, suggesting one or more distinct HA-binding regions on PEDF. The HA-binding region was examined by site-directed mutagenesis.

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Sequence and expression analysis of bovine pigment epithelium-derived factor

Cited by 28 publications

References 24 publications

Pigment epithelium-derived factor in the monkey retinal pigment epithelium and interphotoreceptor matrix: apical secretion and distribution

Pigment epithelium-derived factor in the monkey retinal pigment epithelium and interphotoreceptor matrix: apical secretion and distribution

Identification of a Lipase-linked Cell Membrane Receptor for Pigment Epithelium-derived Factor

Pigment Epithelium-derived Factor Binds to Hyaluronan

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