Laminin receptor involvement in the anti-angiogenic activity of pigment epithelium-derived factor.
Pigment epithelium-derived factor (PEDF) is a multifunctional protein with neurotrophic, anti-oxidative, and anti-inflammatory properties. It is also one of the most potent endogenous inhibitors of angiogenesis, playing an important role in restricting tumor growth, invasion, and metastasis. Studies show that PEDF binds to cell surface proteins, but little is known about how it exerts its effects. Recently, research identified phospholipase A(2)/nutrin/patatin-like phospholipase domain-containing 2 as one PEDF receptor. To identify other receptors, we performed yeast two-hybrid screening using PEDF as bait and discovered that the non-integrin 37/67-kDa laminin receptor (LR) is another PEDF receptor. Co-immunoprecipitation, His tag pulldown, and surface plasmon resonance assays confirmed the interaction between PEDF and LR. Using the yeast two-hybrid method, we further restricted the LR-interacting domain on PEDF to a 34-amino acid (aa) peptide (aa 44-77) and the PEDF-interacting domain on LR to a 91-aa fragment (aa 120-210). A 25-mer peptide named P46 (aa 46-70), derived from 34-mer, interacts with LR in surface plasmon resonance assays and binds to endothelial cell (EC) membranes. This peptide induces EC apoptosis and inhibits EC migration, tube-like network formation in vitro, and retinal angiogenesis ex vivo, like PEDF. Our results suggest that LR is a real PEDF receptor that mediates PEDF angiogenesis inhibition.
Bernard
Adrien
A
Université Pierre et Marie Curie, Univerisité Paris 06, UR4, Aging, Stress and Inflammation and Institut Fédératif de Recherche 83, 75252 Paris, France.
Gao-Li
Jacqueline
J
Franco
Claudio-Areias
CA
Bouceba
Tahar
T
Huet
Alexis
A
Li
Zhenlin
Z
D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D053148:Caspase 3 / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000502:physiology*; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D017474:Receptors, Laminin / Q000235:genetics / Q000378:metabolism*; D015843:Serpins / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D000099174:Pigment Epithelium-Derived Factor
D020533:Angiogenesis Inhibitors; D053148:Caspase 3; D005136:Eye Proteins; D009414:Nerve Growth Factors; D010446:Peptide Fragments; D017474:Receptors, Laminin; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor