A novel interaction between perlecan protein core and progranulin: potential effects on tumor growth.
In an in vivo search of novel partners for perlecan, a major heparan sulfate proteoglycan of basement membranes and cell surfaces, we identified progranulin, a secreted growth factor, as a strong interacting protein. Unambiguous interaction, first observed with the yeast two-hybrid system, was corroborated by co-immunoprecipitation studies using cell-free transcription/translation and transient cell transfection assays. The interaction of progranulin with perlecan domain V involved the first two laminin- and epidermal growth factor-like repeats. Within progranulin, the subdomains interacting most with perlecan harbored granulins F and B. Kinetics analysis of the interaction using surface plasmon resonance showed a saturable binding of relative low affinity (KD approximately 1 microM). These results were supported by significant expression overlap of these two proteins in a series of ovarian tumor tissue microarrays. Progranulin was present within proliferating blood vessels of ovarian carcinomas and perivascular matrices, with a distribution similar to perlecan. Notably, both progranulin and domain V stimulated the growth of adrenal carcinoma cells. However, when used together in equimolar amounts, the two proteins counteracted each other's activity. Thus, progranulin/perlecan interaction could contribute to a fine regulation of tumor angiogenesis and could ultimately affect cancer growth.
Gonzalez
Eva M
EM
Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Mongiat
Maurizio
M
Slater
Simon J
SJ
Baffa
Raffaele
R
Iozzo
Renato V
RV
D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D002455:Cell Division; D002460:Cell Line; D002474:Cell-Free System; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006133:Growth Substances / Q000737:chemistry* / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D009369:Neoplasms / Q000378:metabolism*; D009389:Neovascularization, Pathologic; D020411:Oligonucleotide Array Sequence Analysis; D011233:Precipitin Tests; D000077153:Progranulins; D011485:Protein Binding; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D020349:Surface Plasmon Resonance; D014162:Transfection; D014407:Tumor Cells, Cultured; D020798:Two-Hybrid System Techniques
D020533:Angiogenesis Inhibitors; D018076:DNA, Complementary; C492314:GRN protein, human; D006133:Growth Substances; D019812:Heparan Sulfate Proteoglycans; D036341:Intercellular Signaling Peptides and Proteins; D000077153:Progranulins; D011994:Recombinant Proteins; C071927:perlecan