TIN-ag-RP, a novel catalytically inactive cathepsin B-related protein with EGF domains, is predominantly expressed in vascular smooth muscle cells.
A human cDNA of 2166 bp encoding a novel cathepsin B-related protein was isolated and characterized. The amino acid sequence of the predicted protein of 467 aa was 46% identical with that of human tubulointerstitial nephritis antigen (TIN-ag), and therefore, the protein was tentatively designated as the TIN-ag-related protein (TIN-ag-RP). The amino acid sequence of TIN-ag-RP is composed of a 21 aa long signal sequence, a 181 aa long N-terminal domain containing two epidermal growth factor-like domains, a follistatin motif, and a 265 aa long cathepsin B-like domain. Interestingly, a serine residue has replaced the active site cysteine residue in the cathepsin B-like domain, resulting in a proteolytically inactive protein. Evolutionary analysis revealed that a distinct family of "TIN-ag-like" proteins had evolved in vertebrates. Northern blot analysis revealed a single TIN-ag-RP transcript of 2.4 kb in various tissues with the highest transcript levels detected in aorta, heart, placenta, skeletal muscle, kidney, and a colorectal adenocarcinoma cell line. Using a polyclonal anti-TIN-ag-RP antibody, TIN-ag-RP expression was predominantly seen in vascular smooth muscle (VSM) cells, but also in cardiac and skeletal muscle cells as well as in kidney. Interestingly, uterine smooth muscle cells completely lacked TIN-ag-RP expression, implying a regulated gene expression. Localization studies in HeLa cells stably transfected with TIN-ag-RP cDNA showed that TIN-ag-RP is glycosylated and actively secreted, a finding in line with its proposed function as a structural or regulatory protein similar to TIN-ag.
Wex
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T
Department of Human Genetics, Mount Sinai School of Medicine, New York, New York 10029, USA.
Lipyansky
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A
Brömme
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NC
Wex
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H
Guan
X Q
XQ
Brömme
D
D
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface; D001483:Base Sequence; D002384:Catalysis; D002401:Cathepsin B / Q000737:chemistry*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D018076:DNA, Complementary / Q000737:chemistry; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000737:chemistry*; D016326:Extracellular Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D054834:Lipocalins; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D009395:Nephritis, Interstitial / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D034501:Telomere-Binding Proteins; D014162:Transfection; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D054834:Lipocalins; D008562:Membrane Glycoproteins; D009124:Muscle Proteins; D011994:Recombinant Proteins; C085434:TINAG protein, human; C423605:TINAGL1 protein, human; C440868:TINF2 protein, human; D034501:Telomere-Binding Proteins; D004815:Epidermal Growth Factor; D002401:Cathepsin B