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Simultaneous isolation of human kidney cathepsins B, H, L and C and their characterisation


, : Simultaneous isolation of human kidney cathepsins B, H, L and C and their characterisation. Journal of Chromatography B Biomedical Applications 681(2): 251-262

A procedure for the simultaneous isolation of four cysteine proteinases, cathepsins B, H, L and C, from human kidney is described. The method includes concentration of the acidified homogenate by ammonium sulphate precipitation. The resuspended and dialysed precipitate was chromatographed on DEAE-cellulose DE-32, to allow separation of cathepsins H and C from cathepsins B and L. The main isoform of cathepsin H was separated from cathepsin C by cation-exchange chromatography on CM-Sephadex C-50. These two enzymes were further purified by covalent chromatography on thiopropyl Sepharose and gel permeation on Sephacryl S-200. The last step allowed separation of cathepsin C and the minor isoform of cathepsin H. Purification of the other two enzymes, cathepsins B and L, was carried out on thiol Sepharose, followed by chromatography on CM-Sepharose C-50. In this step, pure cathepsin L was obtained, while two isoforms of cathepsin B had to be finally purified on Sephacryl S-200 columns. The purity of each enzyme was analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis, isoelectric focusing on polyacrylamide gels and N-terminal sequencing. The activities of the purified cathepsins B, H and L were determined in terms of k-cat/K-M for three substrates, Z-Phe-Arg-MCA, Z-Arg-Arg-MCA and Arg-MCA. The method produced 25 mg of cathepsin B, 6.5 mg of cathepsin H, 1.5 mg of cathepsin L and 3.8 mg of cathepsin C from 3.5 kg of human kidney.

US$19.90

PMID: 8811434

DOI: 10.1016/0378-4347(95)00555-2


Other references

Matthews, L.J., 1960: Weed identification and control. Buttercups. An identification key is given for Ranunculus spp. parviflorus, acris, repens, sardous, flammula, rivularis and sceleratus. The last 3 are recognised as poisonous. Recommendations for control are included.

Emmerich, B., 1984: Therapy of malignant gammopathies. Deutsche Medizinische Wochenschrift 109(1): 26-30

Gelabert, R.; Solis, L., 1994: The selection of food particle size by Artemia from Guantanamo, Cuba La seleccion del tamano de particulas alimenticias por la Artemia de Guantanamo, Cuba. Revista de Investigaciones Marinas, 152: 141-145

Broughton, G.; Ilagan, M.; Jaffin, J., 1998: Picture archiving communication system does not decrease the number of radiographs needed to evaluate the trauma patient. Background. Picture Archiving Communication System (PACS) is a sophisticated software and hardware package that enables clinicians to retrieve, review, and digitally manipulate radiographs from computer workstations throughout the hospital. PACS w...

Liang, L.; Liu, Z-Xiao.; Pan, S-Cheng.; Zhang, X-Yan.; Bai, Z-Qing.; Wang, C-Hua.; Yang, M-Hua., 2010: Diagnosis of chronic enteritis of alpine musk deer (Moschus chrysogaster) based on visible-near infrared reflectance spectra of feces. A new method was put forward to diagnose chronic enteritis of alpine musk deer (Moschus chrysogaster) by visible-near infrared reflectance spectra of feces. A total of 125 feces samples, including 70 samples from healthy individuals (healthy sampl...

Barut, B.A.; Zon, L.I.; Cochran, M.K.; Paul, S.R.; Chauhan, D.; Mohrbacher, A.; Fingeroth, J.; Anderson, K.C., 1992: Role of interleukin 6 in the growth of myeloma-derived cell lines. The role of interleukin 6 (IL-6) in the growth of five multiple myeloma-derived cell lines was characterized. The U266 and RPMI 8226 cells lines demonstrated increased DNA synthesis when cultured with exogenous IL-6, expressed IL-6 cell surface re...

Wang, C.; Shimizu Okabe, C.; Watanabe, K.; Okabe, A.; Matsuzaki, H.; Ogawa, T.; Mori, N.; Fukuda, A.; Sato, K., 2002: Developmental changes in KCC1, KCC2, and NKCC1 mRNA expressions in the rat brain. We investigated the expressions of KCC1, KCC2 and NKCC1 mRNAs in the developing rat brain. The neuroepithelium showed abundant KCC1 and NKCC1 mRNA expressions, while KCC2 mRNA was not detected there. In contrast, KCC2 mRNA was preferentially expre...

Wihlborg, O., 1986: The effect of a change in management of displaced tibial shaft fractures. Rigid fixation versus conservative treatment. Helvetica Chirurgica Acta 53(1-2): 191-199

South, M.A., 1978: Conversations on allergy and immunology: does immunity make you sick?. Cutis 21(1): 23, 30-1

Dougherty, B.J.; Fields, D.P.; Mitchell, G.S., 2016: Mammalian target of rapamycin is required for phrenic long-term facilitation following severe but not moderate acute intermittent hypoxia. Phrenic long-term facilitation (pLTF) is a persistent increase in phrenic nerve activity after acute intermittent hypoxia (AIH). Distinct cell-signaling cascades give rise to pLTF depending on the severity of hypoxemia within hypoxic episodes. Mod...