Human glutamate carboxypeptidase 2 (GCP2) from the M28B metalloprotease group is an important target for therapy in neurological disorders and an established tumor marker. However, its physiological functions remain unclear. To better understand general roles, we used the model organism Caenorhabditis elegans to genetically manipulate its three existing orthologous genes and evaluate the impact on worm physiology. The results of gene knockout studies showed that C. elegans GCP2 orthologs affect the pharyngeal physiology, reproduction, and structural integrity of the organism. Promoter-driven GFP expression revealed distinct localization for each of the three gene paralogs, with gcp-2.1 being most abundant in muscles, intestine, and pharyngeal interneurons, gcp-2.2 restricted to the phasmid neurons, and gcp-2.3 located in the excretory cell. The present study provides new insight into the unique phenotypic effects of GCP2 gene knockouts in C. elegans, and the specific tissue localizations. We believe that elucidation of particular roles in a non-mammalian organism can help to explain important questions linked to physiology of this protease group and in extension to human GCP2 involvement in pathophysiological processes.

Department of Cell Biology Faculty of Science Charles University Vinicna 7 Prague 2 128 00 Czech Republic

Department of Chemistry Faculty of Agrobiology Food and Natural Resources Czech University of Life Sciences Prague Kamycka 129 Prague 165 00 Czech Republic

Department of Parasitology Faculty of Science Charles University Vinicna 7 Prague 2 128 00 Czech Republic

Department of Pharmaceutical Chemistry University of California San Francisco 1700 4th Street CA 94143 USA

Department of Physiology University of California San Francisco 600 16th Street CA 94143 U S A

Department of Zoology and Fisheries Center of Infectious Animal Diseases Faculty of Agrobiology Food and Natural Resources Czech University of Life Sciences Prague Kamycka 129 Prague 165 00 Czech Republic

Faculty of Environmental Sciences Center of Infectious Animal Diseases Czech University of Life Sciences Prague Kamycka 129 Prague 165 00 Czech Republic

Institute of Organic Chemistry and Biochemistry The Czech Academy of Sciences Flemingovo n 2 Prague 160 00 Czech Republic

Laboratory of Structural Biology Institute of Biotechnology of the Czech Academy of Sciences BIOCEV Prumyslova 595 Vestec 252 50 Czech Republic

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