Human procathepsin D: three-dimensional model and isolation.
Human procathepsin D was isolated from medium of human breast cancer cell line ZR-75-1 potentiated with estrogen. The isolation involved both immunoaffinity chromatography and ion-exchange chromatography. The affinity chromatography employed polyclonal antibodies raised against a synthetic activation peptide of human cathepsin D. We have started preliminary crystallization trials using the isolated material. A model of human procathepsin D was also built using coordinates of human cathepsin D and pig pepsinogen. The model aids understanding of multiple roles played by activation peptides of aspartic proteinases and will be used as a starting model for molecular replacement.
Oklahoma Medical Research Foundation, Oklahoma City 73104, USA.
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Koelsch
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Metcalf
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Vetvicka
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Fusek
D000595:Amino Acid Sequence; D000818:Animals; D001943:Breast Neoplasms; D002402:Cathepsin D / Q000737:chemistry* / Q000302:isolation & purification*; D002460:Cell Line; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D018360:Crystallography, X-Ray; D004792:Enzyme Precursors / Q000737:chemistry* / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010435:Pepsinogens / Q000737:chemistry; D011487:Protein Conformation; D013552:Swine; D014407:Tumor Cells, Cultured
D004792:Enzyme Precursors; D010435:Pepsinogens; C058345:procathepsin D; D002402:Cathepsin D