| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Investigative Ophthalmology & Visual Science, Vol 37, 1984-1993, Copyright © 1996 by Association for Research in Vision and Ophthalmology
ARTICLES AND REPORTS |
YQ Wu and SP Becerra
Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892- 2740, USA.
PURPOSE: Experiments were designed to identify proteolytic activities that cleave pigment epithelium-derived factor (PEDF), a member of the serpin (serine protease inhibitor) family. METHODS: Proteins in vitreous humor from bovine eyes were analyzed by Western blot with antiserum to human recombinant PEDF protein. Protein fractionation was by ammonium sulfate saturation and by S-Sepharose column chromatography. Proteolytic activities were determined by gelatin zymography and by solution assays against PEDF or chromogenic peptide substrates. RESULTS: PEDF protein was identified and purified to near homogeneity from vitreous humor of bovine eyes. Limited proteolysis showed that the vitreal protein has a protease-sensitive region at its serpin-exposed peptide loop. Proteolytic activities that cleave the PEDF 49.5 kDa-polypeptide were identified only when proteins from these extracts were separated by 45% to 70% ammonium sulfate fractionation (P70). The degradation product had an apparent molecular weight of 46 kDa. This result is consistent with cleavage at the serpin-exposed loop. The PEDF-cleavage activity in P70 was inhibited specifically by the serine protease inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF), but not by aprotinin, EDTA, or pepstatin. The vitreal P70 extracts contained 49- and 53-kDa gelatinolytic activities that also were inhibited by AEBSF and not by EDTA, aprotinin, or pepstatin. The PEDF-cleavage activity did not hydrolyze substrates for thrombin, factor Xa, alpha-chymotrypsin, trypsin, or plasmin, nor did it immunoreact with antibody to urokinase plasminogen activator. CONCLUSIONS: These data indicated that vitreous has a serine- proteolytic activity associated with a novel 49/53-kDa enzyme that cleaves the PEDF protein in a serpinase fashion. In addition to cleavage in vitro, these proteases might play a role in modulating PEDF in vivo.
This article has been cited by other articles:
|
|
 |
|
  S. P. Becerra, L. A. Perez-Mediavilla, J. E. Weldon, S. Locatelli-Hoops, P. Senanayake, L. Notari, V. Notario, and J. G. Hollyfield Pigment Epithelium-derived Factor Binds to Hyaluronan: MAPPING OF A HYALURONAN BINDING SITE J. Biol. Chem., November 28, 2008; 283(48): 33310 - 33320. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Valnickova, S. V. Petersen, S. B. Nielsen, D. E. Otzen, and J. J. Enghild Heparin Binding Induces a Conformational Change in Pigment Epithelium-derived Factor J. Biol. Chem., March 2, 2007; 282(9): 6661 - 6667. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Amaral, R. N. Fariss, M. M. Campos, W. G. Robison Jr, H. Kim, R. Lutz, and S. P. Becerra Transscleral-RPE Permeability of PEDF and Ovalbumin Proteins: Implications for Subconjunctival Protein Delivery Invest. Ophthalmol. Vis. Sci., December 1, 2005; 46(12): 4383 - 4392. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Notari, A. Miller, A. Martinez, J. Amaral, M. Ju, G. Robinson, L. E. H. Smith, and S. P. Becerra Pigment Epithelium-Derived Factor Is a Substrate for Matrix Metalloproteinase Type 2 and Type 9: Implications for Downregulation in Hypoxia Invest. Ophthalmol. Vis. Sci., August 1, 2005; 46(8): 2736 - 2747. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Yamane, A. Minamoto, H. Yamashita, H. Takamura, Y. Miyamoto-Myoken, K. Yoshizato, T. Nabetani, A. Tsugita, and H. K. Mishima Proteome Analysis of Human Vitreous Proteins Mol. Cell. Proteomics, November 1, 2003; 2(11): 1177 - 1187. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Meyer, L. Notari, and S. P. Becerra Mapping the Type I Collagen-binding Site on Pigment Epithelium-derived Factor. IMPLICATIONS FOR ITS ANTIANGIOGENIC ACTIVITY J. Biol. Chem., November 15, 2002; 277(47): 45400 - 45407. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. M. Bilak, S. P. Becerra, A. M. Vincent, B. H. Moss, M. S. Aymerich, and R. W. Kuncl Identification of the Neuroprotective Molecular Region of Pigment Epithelium-Derived Factor and Its Binding Sites on Motor Neurons J. Neurosci., November 1, 2002; 22(21): 9378 - 9386. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Z. Renno, A. I. Youssri, N. Michaud, E. S. Gragoudas, and J. W. Miller Expression of Pigment Epithelium-Derived Factor in Experimental Choroidal Neovascularization Invest. Ophthalmol. Vis. Sci., May 1, 2002; 43(5): 1574 - 1580. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Ogata, M. Wada, T. Otsuji, N. Jo, J. Tombran-Tink, and M. Matsumura Expression of Pigment Epithelium-Derived Factor in Normal Adult Rat Eye and Experimental Choroidal Neovascularization Invest. Ophthalmol. Vis. Sci., April 1, 2002; 43(4): 1168 - 1175. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. J. Duh, H. S. Yang, I. Suzuma, M. Miyagi, E. Youngman, K. Mori, M. Katai, L. Yan, K. Suzuma, K. West, et al. Pigment Epithelium-Derived Factor Suppresses Ischemia-Induced Retinal Neovascularization and VEGF-Induced Migration and Growth Invest. Ophthalmol. Vis. Sci., March 1, 2002; 43(3): 821 - 829. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. S. Aymerich, E. M. Alberdi, A. Martinez, and S. P. Becerra Evidence for Pigment Epithelium-Derived Factor Receptors in the Neural Retina Invest. Ophthalmol. Vis. Sci., December 1, 2001; 42(13): 3287 - 3293. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Simonovic, P. G. W. Gettins, and K. Volz Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor PNAS, September 13, 2001; (2001) 211268598. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Alberdi, M. S. Aymerich, and S. P. Becerra Binding of Pigment Epithelium-derived Factor (PEDF) to Retinoblastoma Cells and Cerebellar Granule Neurons. EVIDENCE FOR A PEDF RECEPTOR J. Biol. Chem., October 29, 1999; 274(44): 31605 - 31612. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. W. Dawson, O. V. Volpert, P. Gillis, S. E. Crawford, H. Xu, W. Benedict, and N. P. Bouck Pigment Epithelium-Derived Factor: A Potent Inhibitor of Angiogenesis Science, July 9, 1999; 285(5425): 245 - 248. [Abstract] [Full Text]
|
|
|
|
|
|
M. Simonovic, P. G. W. Gettins, and K. Volz Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factor PNAS, September 25, 2001; 98(20): 11131 - 11135. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
