Recombinant Human Cathepsin L Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human Cathepsin L Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008). The specific activity is >25,000 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Cathepsin L protein Glu113-Val333 & Ala114-Val333, both with a C-terminal 6-His tag
• Accession #: P07711
• N-terminal Sequence: Glu113 & Ala114
• Structure / Form: Mature form
• Protein/Peptide Type: Recombinant Enzymes
• Gene: CTSL
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 26 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 36 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  6 months from date of receipt, -20 to -70 degreesC as supplied. 3 months, -20 to -70 degreesC under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 50 mM MES, 5 mM DTT, 1 mM EDTA, 0.005% (w/v) Brij-35, pH 6.0
  • Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
  • Fluorogenic Peptide Substrate VII: Z-Leu-Arg-AMC (Catalog # ES008)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCathepsin L to 40 µg/mL in Assay Buffer.
  2. Incubate diluted rhCathepsin L on ice for 15 minutes.
  3. Dilute incubated 40 µg/mL rhCathepsin L to 0.02 ng/µL in Assay Buffer.
  4. Dilute Substrate to 80 µM in Assay Buffer.
  5. Load 50 µL of 0.02 ng/µL rhCathepsin L into a black well plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 µM Substrate without any rhCathepsin L.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)) / (amount of enzyme (µg))

  *Adjusted for Substrate Blank

  **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).

  Per Well:
  • rhCathepsin L: 0.001 µg
  • Substrate: 40 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Cathepsin L Protein, CF

• Cathepsin L
• cathepsin L1
• CATL
• CTSL
• CTSL1
• CTSLEC 3.4.22.15
• EC 3.4.22
• FLJ31037
• Major excreted protein
• MEP

Background

Cathepsin L is a lysosomal cysteine protease expressed in most eukaryotic cells. Cathepsin L is known to hydrolyze a number of proteins, including the proform of urokinase-type plasminogen activator, which is activated by Cathepsin L cleavage (1). Cathepsin L has also been shown to proteolytically inactivate alpha 1 -antitrypsin and secretory leucoprotease inhibitor, two major protease inhibitors of the respiratory tract (2). These observations, combined with the demonstration of increased Cathepsin L activity in the epithelial lining fluid of the lungs of emphysema patients, have led to the suggestion that the enzyme may be involved in the progression of this disease. Cathepsin L has also been identified as a major excreted protein of transformed fibroblasts, indicating the enzyme could be involved in malignant tumor growth (3). Human Cathepsin L activity is greatest under mildly acidic conditions, from pH 4.5 - 6.5. The stability of the enzyme decreases at higher pH values.

1. Goretzki, L. et al. (1992) FEBS Lett. 297:112.
2. Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
3. Gottesman, M.M. and F. Cabral (1981) Biochemistry 20:1659.

Publications for Cathepsin L (952-CY)(30)

Publications using 952-CY

• Chazot, A;Zimberger, C;Feracci, M;Moussa, A;Good, S;Sommadossi, JP;Alvarez, K;Ferron, F;Canard, B; The activation cascade of the broad-spectrum antiviral bemnifosbuvir characterized at atomic resolution PLoS biology 2024-08-01 [PMID: 39190717] (Bioassay, N/A)
• Sutton, VR;Watt, SV;Akhlaghi, H;Cipolla, DC;Chen, KJ;LaSala, D;McDonald, PP;Beavis, PA;Munoz, I;Hodel, AW;Noori, T;Voskoboinik, I;Trapani, JA; Pharmacologic inhibition of dipeptidyl peptidase 1 (cathepsin C) does not block in vitro granzyme-mediated target cell killing by CD8 T or NK cells Frontiers in pharmacology 2024-07-03 [PMID: 39021839] (Bioassay, N/A)
• Heggelund, JE;Das, S;Stamnaes, J;Iversen, R;Sollid, LM; Autoantibody binding and unique enzyme-substrate intermediate conformation of human transglutaminase 3 Nature communications 2023-10-05 [PMID: 37798283] (Bioassay, N/A)
• CJ Nirschl, HR Brodkin, C Domonkos, CJ Dwyer, DJ Hicklin, N Ismail, C Seidel-Dug, P Steiner, Z Steuert, JM Sullivan, WM Winston, A Salmeron mWTX-330, an IL 12 INDUKINE Molecule, Activates and Reshapes Tumor-infiltrating CD8+ T and NK Cells to Generate Antitumor Immunity Cancer Immunology Research, 2023-04-19;0(0):. 2023-04-19 [PMID: 37074216] (Bioassay, Human)
• CJ Nirschl, HR Brodkin, C Domonkos, CJ Dwyer, DJ Hicklin, N Ismail, C Seidel-Dug, P Steiner, Z Steuert, JM Sullivan, WM Winston, A Salmeron mWTX-330, an IL 12 INDUKINE Molecule, Activates and Reshapes Tumor-infiltrating CD8+ T and NK Cells to Generate Antitumor Immunity Cancer Immunology Research, 2023-04-19;0(0):. 2023-04-19 [PMID: 37074216] (Bioassay)
• S Kim, KH Lee, HJ Choi, E Kim, S Kang, M Han, HJ Jeon, MY Yun, GY Song, HJ Lee Hederacolchiside A1 Suppresses Autophagy by Inhibiting Cathepsin C and Reduces the Growth of Colon Cancer Cancers, 2023-02-16;15(4):. 2023-02-16 [PMID: 36831614] (Bioassay, N/A)
• AV Luebben, D Bender, S Becker, LM Crowther, I Erven, K Hofmann, J Söding, H Klemp, C Bellotti, A Stäuble, T Qiu, RS Kathayat, BC Dickinson, J Gärtner, GM Sheldrick, R Krätzner, R Steinfeld Cln5 represents a new type of cysteine-based S-depalmitoylase linked to neurodegeneration Science Advances, 2022-04-15;8(15):eabj8633. 2022-04-15 [PMID: 35427157] (Bioassay, N/A)
• T Shapira, IA Monreal, SP Dion, DW Buchholz, B Imbiakha, AD Olmstead, M Jager, A Désilets, G Gao, M Martins, T Vandal, CAH Thompson, A Chin, WD Rees, T Steiner, IR Nabi, E Marsault, J Sahler, DG Diel, GR Van de Wal, A August, GR Whittaker, PL Boudreault, R Leduc, HC Aguilar, F Jean A TMPRSS2 inhibitor acts as a pan-SARS-CoV-2 prophylactic and therapeutic Nature, 2022-03-28;0(0):. 2022-03-28 [PMID: 35344983] (Bioassay, N/A)
• W Deng, Y Bai, F Deng, Y Pan, S Mei, Z Zheng, R Min, Z Wu, W Li, R Miao, Z Zhang, TS Kupper, J Lieberman, X Liu Streptococcal pyrogenic exotoxin B cleaves GSDMA and triggers pyroptosis Nature, 2022-02-02;0(0):. 2022-02-02 [PMID: 35110732] (Bioassay, Human)
• DDC Linhares, F Faria, RT Kodama, AMXP Amorim, FCV Portaro, D Trevisan-S, KF Ferraz, AM Chudzinski Novel Cysteine Protease Inhibitor Derived from the Haementeria vizottoi Leech: Recombinant Expression, Purification, and Characterization Toxins, 2021-12-02;13(12):. 2021-12-02 [PMID: 34941695] (Bioassay, Haementeria vizzotoi)
• J Li, S Liu, J Shi, HJ Zhu Activation of Tenofovir Alafenamide and Sofosbuvir in the Human Lung and Its Implications in the Development of Nucleoside/Nucleotide Prodrugs for Treating SARS-CoV-2 Pulmonary Infection Pharmaceutics, 2021-10-11;13(10):. 2021-10-11 [PMID: 34683949] (Enzyme Activity, Human)
• S Kim, SI Lee, N Kim, M Joo, KH Lee, MW Lee, HJ Jeon, H Ryu, JM Kim, JY Sul, GY Song, JY Kim, HJ Lee Decursin inhibits cell growth and autophagic flux in gastric cancer via suppression of cathepsin C American journal of cancer research, 2021-04-15;11(4):1304-1320. 2021-04-15 [PMID: 33948359] (Bioassay, Human)
• E Mittler, T Alkutkar, RK Jangra, K Chandran Direct Intracellular Visualization of Ebola Virus-Receptor Interaction by In Situ Proximity Ligation MBio, 2021-01-12;12(1):. 2021-01-12 [PMID: 33436438] (Bioassay, Human)
• RA Høglund, SB Torsetnes, A Lossius, B Bogen, EJ Homan, R Bremel, T Holmøy Human Cysteine Cathepsins Degrade Immunoglobulin G In Vitro in a Predictable Manner Int J Mol Sci, 2019-09-29;20(19):. 2019-09-29 [PMID: 31569504] (Bioassay, Human)
• D Dana, J Garcia, AI Bhuiyan, P Rathod, L Joo, DA Novoa, S Paroly, KR Fath, EJ Chang, SK Pathak Cell penetrable, clickable and tagless activity-based probe of human cathepsin L Bioorg. Chem., 2019-02-14;85(0):505-514. 2019-02-14 [PMID: 30802807] (Click Chemistry, Human)
• R Blau, Y Epshtein, E Pisarevsky, G Tiram, S Israeli Da, E Yeini, A Krivitsky, A Eldar-Booc, D Ben-Shusha, H Gibori, A Scomparin, O Green, Y Ben-Nun, E Merquiol, H Doron, G Blum, N Erez, R Grossman, Z Ram, D Shabat, R Satchi-Fai Image-guided surgery using near-infrared Turn-ON fluorescent nanoprobes for precise detection of tumor margins Theranostics, 2018-05-24;8(13):3437-3460. 2018-05-24 [PMID: 30026858] (Bioassay, N/A)
• JB Schaal, DQ Tran, A Subramania, R Patel, T Laragione, KD Roberts, K Trinh, P Tongaonkar, PA Tran, D Minond, GB Fields, P Beringer, AJ Ouellette, PS Gulko, ME Selsted Suppression and resolution of autoimmune arthritis by rhesus ?-defensin-1, an immunomodulatory macrocyclic peptide PLoS ONE, 2017-11-16;12(11):e0187868. 2017-11-16 [PMID: 29145473] (Bioassay)
• CW Lee, JN Stankowski, J Chew, CN Cook, YW Lam, S Almeida, Y Carlomagno, KF Lau, M Prudencio, FB Gao, M Bogyo, DW Dickson, L Petrucelli The lysosomal protein cathepsin L is a progranulin protease Mol Neurodegener, 2017-07-25;12(1):55. 2017-07-25 [PMID: 28743268] (Bioassay, Human)
• X Zhao, KA Howell, S He, JM Brannan, AZ Wec, E Davidson, HL Turner, CI Chiang, L Lei, JM Fels, H Vu, S Shulenin, AN Turonis, AI Kuehne, G Liu, M Ta, Y Wang, C Sundling, Y Xiao, JS Spence, BJ Doranz, FW Holtsberg, AB Ward, K Chandran, JM Dye, X Qiu, Y Li, MJ Aman Immunization-Elicited Broadly Protective Antibody Reveals Ebolavirus Fusion Loop as a Site of Vulnerability Cell, 2017-05-18;169(5):891-904.e15. 2017-05-18 [PMID: 28525756] (Bioassay, Human)
• AK Piper, SE Ross, GM Redpath, FA Lemckert, N Woolger, A Bournazos, PA Greer, RB Sutton, ST Cooper Enzymatic cleavage of myoferlin releases a dual C2-domain module linked to ERK signalling Cell. Signal, 2017-02-10;33(0):30-40. 2017-02-10 [PMID: 28192161] (Bioassay, Human)
• Shivanna V, Kim Y, Chang K Endosomal acidification and cathepsin L activity is required for calicivirus replication. Virology, 2014-08-09;464(0):287-95. 2014-08-09 [PMID: 25108379] (Bioassay, Porcine)
• Jiang H, Cheng X, Shi G, Hu L, Inoue A, Yamamura Y, Wu H, Takeshita K, Li X, Huang Z, Song H, Asai M, Hao C, Unno K, Koike T, Oshida Y, Okumura K, Murohara T, Kuzuya M Cathepsin K-mediated Notch1 activation contributes to neovascularization in response to hypoxia. Nat Commun, 2014-06-04;5(0):3838. 2014-06-04 [PMID: 24894568] (Bioassay, Mouse)
• Guery L, Benikhlef N, Gautier T, Paul C, Jego G, Dufour E, Jacquel A, Cally R, Manoury B, Vanden Berghe T, Vandenabeele P, Droin N, Solary E Fine-tuning nucleophosmin in macrophage differentiation and activation. Blood, 2011-08-29;118(17):4694-704. 2011-08-29 [PMID: 21876121] (Enzyme Assay, Human)
• Zou F, Schafer N, Palesch D, Brucken R, Beck A, Sienczyk M, Kalbacher H, Sun Z, Boehm BO, Burster T Regulation of cathepsin g reduces the activation of proinsulin-reactive T cells from type 1 diabetes patients. PLoS ONE, 2011-08-05;6(8):e22815. 2011-08-05 [PMID: 21850236] (Enzyme Assay, Human)
• Gonzalez RM, Seurynck-Servoss SL, Crowley SA Development and validation of sandwich ELISA microarrays with minimal assay interference. J. Proteome Res., 2008-04-19;7(6):2406-14. 2008-04-19 [PMID: 18422355] (ELISA (Standard))
• Hamilton G, Colbert JD, Schuettelkopf AW, Watts C Cystatin F is a cathepsin C-directed protease inhibitor regulated by proteolysis. EMBO J., 2008-02-06;27(3):499-508. 2008-02-06 [PMID: 18256700] (Enzyme Assay)
• Burster T, Marin-Esteban V, Boehm BO, Dunn S, Rotzschke O, Falk K, Weber E, Verhelst SH, Kalbacher H, Driessen C Design of protease-resistant myelin basic protein-derived peptides by cleavage site directed amino acid substitutions. Biochem. Pharmacol., 2007-08-02;74(10):1514-23. 2007-08-02 [PMID: 17803968] (Enzyme Assay, Human)
• Methot N, Rubin J, Guay D, Beaulieu C, Ethier D, Reddy TJ, Riendeau D, Percival MD Inhibition of the activation of multiple serine proteases with a cathepsin C inhibitor requires sustained exposure to prevent pro-enzyme processing. J. Biol. Chem., 2007-05-29;282(29):20836-46. 2007-05-29 [PMID: 17535802] (Enzyme Assay)
• Cheng T, Hitomi K, van Vlijmen-Willems IM, de Jongh GJ, Yamamoto K, Nishi K, Watts C, Reinheckel T, Schalkwijk J, Zeeuwen PL Cystatin M/E is a high affinity inhibitor of cathepsin V and cathepsin L by a reactive site that is distinct from the legumain-binding site. A novel clue for the role of cystatin M/E in epidermal cornification. J. Biol. Chem., 2006-03-24;281(23):15893-9. 2006-03-24 [PMID: 16565075] (Enzyme Assay, Human)
• Gocheva V, Zeng W, Ke D, Klimstra D, Reinheckel T, Peters C, Hanahan D, Joyce JA Distinct roles for cysteine cathepsin genes in multistage tumorigenesis. Genes Dev., 2006-02-15;20(5):543-56. 2006-02-15 [PMID: 16481467] (Enzyme Assay, Human)

Bioinformatics

• Gene Symbol: CTSL
• Uniprot:
  • P07711()