Tissue expression of the novel serine carboxypeptidase Scpep1.
We previously identified a novel gene designated retinoid-inducible serine carboxypeptidase (RISC or Scpep1). Here we characterize a polyclonal antibody raised to Scpep1 and assess its localization in mouse cells and tissues. Western blot analysis revealed an immunospecific approximately 35-kDa protein corresponding to endogenous Scpep1. This protein is smaller than the predicted approximately 51-kDa, suggesting that Scpep1 is proteolytically cleaved to a mature enzyme. Immunohistochemical studies demonstrate Scpep1 expression in embryonic heart and vasculature as well as in adult aortic smooth muscle cells and endothelial cells. Scpep1 displays a broad expression pattern in adult tissues with detectable levels in epithelia of digestive tract and urinary bladder, islet of Langerhans, type II alveolar cells and macrophages of lung, macrophage-like cells of lymph nodes and spleen, Leydig cells of testis, and nerve fibers in brain and ganglia. Consistent with previous mRNA studies in kidney, Scpep1 protein is restricted to proximal convoluted tubular epithelium (PCT). Immunoelectron microscopy shows enriched Scpep1 within lysosomes of the PCT, and immunofluorescence microscopy colocalizes Scpep1 with lysosomal-associated membrane protein-2. These results suggest that Scpep1 is a widely distributed lysosomal protease requiring proteolytic cleavage for activity. The highly specific Scpep1 antibody characterized herein provides a necessary reagent for elucidating Scpep1 function.
Lee
Ting-Hein D
TH
Cardiovascular Research Institute, University of Rochester School of Medicine, 601 Elmwood Avenue, Box 679, Rochester, NY 14642, USA.
Streb
Jeffrey W
JW
Georger
Mary A
MA
Miano
Joseph M
JM
D000818:Animals; D000906:Antibodies; D002268:Carboxypeptidases / Q000096:biosynthesis* / Q000276:immunology; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry; D007687:Kidney Tubules, Proximal / Q000201:enzymology; D052119:Lysosomal-Associated Membrane Protein 2; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D009928:Organ Specificity
D000906:Antibodies; C497811:LAMP2 protein, human; D052119:Lysosomal-Associated Membrane Protein 2; D051907:Lysosomal Membrane Proteins; D002268:Carboxypeptidases; C436327:SCPEP1 protein, human; C486671:Scpep1 protein, mouse