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Characterization of GPR56 protein and its suppressed expression in human pancreatic cancer cells

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Abstract

GPR56 is an atypical G protein-coupled receptor (GPCR) with an unusually large N-terminal extracellular region, which contains a long Ser/Thr-rich region forming a mucin-like stalk and due to this feature, GPR56 is thought to be an adhesion GPCR. Recent studies demonstrate that GPR56 plays a role in brain development and tumorigenesis. Here, we report that human GPR56 undergoes GPS (GPCR proteolytic site)-mediated protein cleavage to generate its extracellular domain as an N-terminal fragment (GPR56-N). We also show that GPR56-N is highly glycosylated with N-linked carbohydrate chains. Mouse Gpr56 is ubiquitously expressed in various tissues, with high levels in kidney and pancreas. GPR56 mRNA is detected in diverse human cancer cells including pancreatic cancer cells PANC-1, Capan-1, and MiaCaPa-2. Interestingly, GPR56 protein is either negligible or undetectable in these pancreatic cancer cells, despite the fact that high levels of GPR56 mRNA are observed. Moreover, we have found that protein levels of GPR56 in pancreatic cancer cells were not affected when cells were treated with a proteasome inhibitor MG132. Taken together, these results define the biochemical properties of GPR56 protein, and suggest that the expression of GPR56 protein is suppressed in human pancreatic cancer cells.

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Acknowledgements

This work was supported in part by the U.S. National Science Foundation EPSCoR Grant # 0132740 to the state of West Virginia and by a pilot grant from the U.S. National Institutes of Health NCRR Grant # 5P20 RR020180 (PI, Richard Niles, Marshall University). We thank Dr. Christopher Stipp (University of Iowa) for kindly providing us the construct pLXIZ-NFLGPR56, Drs. Andrea McClatchey and Ichiko Saotome (Harvard University) for generously providing us the protein samples from wild-type and ezrin conditional knockout mouse intestines.

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Authors and Affiliations

  1. Biomedical Science Graduate Program, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV, 25704, USA

    Yue Huang & Guo-Zhang Zhu

  2. Department of Biological Sciences, Marshall University, Huntington, WV, 25755, USA

    Jun Fan & Guo-Zhang Zhu

  3. Columbus Children’s Research Institute, 700 Children’s Dr, Columbus, OH, 43205, USA

    Jing Yang

  4. Department of Pediatrics, the Ohio State University, 700 Children’s Dr, Columbus, OH, 43205, USA

    Jing Yang

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  1. Yue Huang
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  2. Jun Fan
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Correspondence to Guo-Zhang Zhu.

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Yue Huang and Jun Fan contributed equally to this work.

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Huang, Y., Fan, J., Yang, J. et al. Characterization of GPR56 protein and its suppressed expression in human pancreatic cancer cells. Mol Cell Biochem 308, 133–139 (2008). https://doi.org/10.1007/s11010-007-9621-4

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  • DOI: https://doi.org/10.1007/s11010-007-9621-4

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