A Novel Interaction between Perlecan Protein Core and Progranulin

Abstract

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 (K D ϳ1 M). 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.

The granulin gene family: from cancer to dementia

“…(92) This interaction has also been implicated in the pathogenesis of arthritis. (93) PGRN binds the ECM proteins perlecan (94) and chondrocyte oligomeric matrix protein (COMP); (95) the perlecan interaction decreases the proliferative activity of PGRN, whereas the COMP interaction enhances it. In both cases PGRN binds its partner weakly, with affinities in the micromolar range, (94,95) and associates with the target protein through an EGF-containing module.…”

“…(93) PGRN binds the ECM proteins perlecan (94) and chondrocyte oligomeric matrix protein (COMP); (95) the perlecan interaction decreases the proliferative activity of PGRN, whereas the COMP interaction enhances it. In both cases PGRN binds its partner weakly, with affinities in the micromolar range, (94,95) and associates with the target protein through an EGF-containing module. PGRN reportedly binds the membrane protein Dlk, (96) which is also rich in EGF modules, and although not yet tested, an affinity for EGF modules may prove a recurrent pattern in PGRN-protein interactions.…”

BMP-1/Tolloid-like Metalloproteases Process Endorepellin, the Angiostatic C-terminal Fragment of Perlecan

“…SDS-PAGE and immunoblottings were as described previously (21). Rabbit polyclonal primary antibodies against endorepellin (22) and procollagen C-proteinase enhancer (23) were used at dilutions of 1:2000 and 1:20,000, respectively. N-terminal sequences were determined by automated Edman degradation at the Harvard University Microchemistry Facility.…”

Perlecan Immunolocalizes to Perichondrial Vessels and Canals in Human Fetal Cartilaginous Primordia in Early Vascular and Matrix Remodeling Events Associated with Diarthrodial Joint Development

“…Perlecan also interacts with type XVIII collagen, and the anti-angiogenic peptide derived from the carboxyl domain of type XVIII collagen, endostatin, which co-localizes with perlecan in a number of human tissues (Miosge et al 2003). Furthermore, perlecan also interacts with progranulin and promotes angiogenesis during tumor development (Gonzalez et al 2003). Perlecan can therefore display both pro-and anti-angiogenic properties, depending on its developmental environment, and undoubtably has important regulatory roles in vascular remodeling (Nakahama et al 2000;Gonzalez et al 2003;Jiang and Couchman 2003).…”