, Volume 71, Issue 5–6, pp 421–432 | Cite as

Identification and characterization of the lamprey cathepsin genes

  • Dong Wang
  • Peng Su
  • Xuefeng Wang
  • Kai Liu
  • Changzhi Li
  • Xingxing Gao
  • Jiali Lu
  • Feng Sun
  • Qingwei LiEmail author
  • Yue PangEmail author
Original Article


Cathepsins are key mammalian proteases that play an important role in the immune response. Several studies have revealed the versatile and critical functions of cathepsins. Here, we obtained ten kinds of cathepsin homologs and identified seven homologs with complete coding sequences. Phylogenetic analysis verified their identities and supported the classification of cathepsins into seven families, which is similar to other vertebrates. Tissue-specific expression analysis showed that all lamprey cathepsins (L-cathepsins) are present in the supraneural body (SB), kidney, gill, intestine, brain, heart, and liver, but their relative abundance varied among tissues. Additionally, we focused on the lamprey cathepsin L (L-cathepsin L) and used recombinant L-cathepsin L protein (rL-cathepsin L) to prepare anti rL-cathepsin L polyclonal antibodies, which were used to detect its distribution in lamprey tissues. The L-cathepsin L protein was primarily detected in the SB, kidney, gill, intestine, brain, and liver via western blot and immunohistochemistry assays. Importantly, quantitative real-time PCR (RT-PCR) revealed that the expression level of L-cathepsins mRNA significantly increased after exposure to three different stimuli (poly I:C, Staphylococcus aureus (S.a) and Vibro anguilarum ( This suggested that L-cathepsins may participate in defense processes. These results revealed that L-cathepsins may play key roles in the immune response to exogenous stimuli. The findings provide important information for future studies aiming to understand the molecular mechanisms underlying the immune response to pathogen invasion in lamprey.


Lamprey Cathepsins Evolution Immune 


Funding information

This work was funded by the Chinese Major State Basic Research Development Program (973 Program; Grant2013CB835304), the Marine Public Welfare Project of the State Oceanic Administration (No.201305016), Chinese National Natural Science Foundation Grants (No. 31170353, No. 31202020, No. 31772884, and No. 31801973), Science and Technology Innovation Fund Research Project (No. 2018J12SN079), and the project of Department of Ocean and Fisheries of Liaoning Province (No. 201805).

Compliance with ethical standards

Competing interests

The authors declare that they have no conflicts of interest.

Supplementary material

251_2019_1117_Fig7_ESM.png (1 mb)
Fig. S1

Fig. 1 Expression of L-cathepsin L recombinant proteins and production, affinity purification and identification of anti- rL-cathepsin L antibodies. a Expression of rL-cathepsin L protein in E. coli BL21 (DE3). Lane 1: total protein of uninduced E. coli harboring pCold I-L-cathepsin L; Lane 2-6: total protein of induced E. coli harboring pCold I-L-cathepsin L after treatment with 0.1 mM, 0.05 mM, 0.15 mM, 0.01 mM and 0.5 mM IPTG for 24 h at 16℃; Lane 7-8: supernatant precipitation from induced E. coli BL21 (DE3) harboring pCold I-L-cathepsin L after treatment with 0.1 mM IPTG for 24 h at 16℃ after sonication. The arrow points to L-cathepsin L. b Purification of rL-cathepsin L protein by HisTrap™ affinity columns. Lane 1: supernatant from induced E. coli BL21 (DE3) harboring pCold I-L-cathepsin L after sonication; Lane 2: flow through; Lane 3: equilibrium; Lane 4-9: elution by iminazole, concentration gradient of 30, 50, 100, 200, 300 and 400 mM; M: protein marker. The arrow points to rL-cathepsin L. c The titer of anti-rL-cathepsin L polyclonal antibody by ELISA assay. The rabbit sera before immunization was used as a control. d Identification of the anti-rL-cathepsin L polyclonal antibody by SDS-PAGE. Lane 1: rabbit anti-rL-cathepsin L antisera; Lane 2: flow through; Lane 3: equilibrium; Lane 4: elution by 0.1 M glycine-HCI (pH 2.7); M: protein marker. The arrow points to the heavy chains and light chains of the antibody (PNG 1.0 MB)

251_2019_1117_MOESM1_ESM.tif (99.6 mb)
High resolution image (TIF 99.6 MB)


  1. Bruchard M, Mignot G, Derangère V, Chalmin F, Chevriaux A, Végran F, Boireau W, Simon B, Ryffel B, Connat JL, Kanellopoulos J, Martin F, Rébé C, Apetoh L, Ghiringhelli F (2013) Chemotherapy-triggered cathepsin B release in myeloid-derived suppressor cells activates the Nlrp3 inflammasome and promotes tumor growth. Nat Med 19(1):57–64CrossRefGoogle Scholar
  2. Caglic D, Pungercar JR, Pejler G, Turk V, Turk B (2007) Glycosaminoglycans facilitate procathepsin B activation through disruption of propeptide-mature enzyme interactions. J Biol Chem 282(45):33076–33085CrossRefGoogle Scholar
  3. Cocchiaro P, De Pasquale V, Della Morte R, Tafuri S, Avallone L, Pizard A, Moles A, Pavone LM (2017) The multifaceted role of the lysosomal protease cathepsins in kidney disease. Front Cell Dev Biol 5:114CrossRefGoogle Scholar
  4. Conus S, Simon HU (2010) Cathepsins and their involvement in immune responses. Swiss Med Wkly 140:w13042Google Scholar
  5. Cook C, Stankowski JN, Carlomagno Y, Stetler C, Petrucelli L (2014) Acetylation: a new key to unlock tau's role in neurodegeneration. Alzheimers Res Ther 6(3):29CrossRefGoogle Scholar
  6. Criscitiello MF, Ohta Y, Graham MD, Eubanks JO, Chen PL, Flajnik MF (2012) Shark class II invariant chain reveals ancient conserved relationships with cathepsins and MHC class II. Dev Comp Immunol 36(3):521–533CrossRefGoogle Scholar
  7. Dennemärker J, Lohmüller T, Mayerle J, Tacke M, Lerch MM, Coussens LM, Peters C, Reinheckel T (2010a) Deficiency for the cysteine protease cathepsin L promotes tumor progression in mouse epidermis. Oncogene 29(11):1611–1621CrossRefGoogle Scholar
  8. Dennemärker J, Lohmüller T, Müller S, Aguilar SV, Tobin DJ, Peters C, Reinheckel T (2010b) Impaired turnover of autophagolysosomes in cathepsin L deficiency. Biol Chem 391(8):913–922CrossRefGoogle Scholar
  9. Dijkstra JM, Yamaguchi (2019) Ancient features of the MHC class II presentation pathway, and a model for the possible origin of MHC molecules. Immunogenetics 71(3):233–249CrossRefGoogle Scholar
  10. Duncan EM, Muratore-Schroeder TL, Cook RG, Garcia BA, Shabanowitz J, Hunt DF, Allis CD (2008) Cathepsin L proteolytically processes histone H3 during mouse embryonic stem cell differentiation. Cell 135(2):284–294CrossRefGoogle Scholar
  11. Godahewa GI, Perera NCN, Lee S, Kim MJ, Lee J (2017) A cysteine protease (cathepsin Z) from disk abalone, Haliotis discus discus: genomic characterization and transcriptional profiling during bacterial infections. Gene 627:500–507CrossRefGoogle Scholar
  12. Goulet B, Baruch A, Moon NS, Poirier M, Sansregret LL, Erickson A, Bogyo M, Nepveu A (2004) A cathepsin L isoform that is devoid of a signal peptide localizes to the nucleus in S phase and processes the CDP/Cux transcription factor. Mol Cell 14(2):207–219CrossRefGoogle Scholar
  13. Gustafsson OS, Collin SP, Kröger RH (2008) Early evolution of multifocal optics for well-focused colour vision in vertebrates. J Exp Biol 211(Pt 10):1559–1564CrossRefGoogle Scholar
  14. Hsing LC, Rydensky AY (2005) The lysosomal cysteine proteases in MHC class II antigen presentation. Immunol Rev 207:229–241CrossRefGoogle Scholar
  15. Ishidoh K, Kominami E (2002) Processing and activation of lysosomal proteinases. Biol Chem 383(12):1827–1831CrossRefGoogle Scholar
  16. Lecaille F, Kaleta J, Brömme D (2002) Human and parasitic papain-like cysteine proteases: their role in physiology and pathology and recent developments in inhibitor design. Chem Rev 102(12):4459–4488CrossRefGoogle Scholar
  17. Nair SV, Del Valle H, Gross PS, Terwilliger DP, Smith LC (2005) Macroarray analysis of coelomocyte gene expression in response to LPS in the sea urchin. Identification of unexpected immune diversity in an invertebrate. Physiol Genomics 22(1):33–47CrossRefGoogle Scholar
  18. Navab R, Pedraza C, Fallavollita L, Wang N, Chevet E, Auguste P, Jenna S, You Z, Bikfalvi A, Hu J, O'Connor R, Erickson A, Mort JS, Brodt P (2008) Loss of responsiveness to IGF-I in cells with reduced cathepsin L expression levels. Oncogene 27(37):4973–4985CrossRefGoogle Scholar
  19. Nikitina N, Bronner-Fraser M, Sauka-Spengler T (2009) The sea lamprey Petromyzon marinus: a model for evolutionary and developmental biology. Cold Spring Harb Protoc 2009(1):pdb.emo113CrossRefGoogle Scholar
  20. Olson OC, Joyce JA (2015) Cysteine cathepsin proteases: regulators of cancer progression and therapeutic response. Nat Rev Cancer 15(12):712–729CrossRefGoogle Scholar
  21. Palermo C, Joyce JA (2008) Cysteine cathepsin proteases as pharmacological targets in cancer. Trends Pharmacol Sci 29(1):22–28CrossRefGoogle Scholar
  22. Pan L, Li Y, Jia L, Qin Y, Qi G, Cheng J, Qi Y, Li H, Du J (2012) Cathepsin S deficiency results in abnormal accumulation of autophagosomes in macrophages and enhances Ang II-induced cardiac inflammation. PLoS One 7(4):e35315CrossRefGoogle Scholar
  23. Pang Y, Wang S, Ba W, Li Q (2015) Cell secretion from the adult lamprey supraneural body tissues possesses cytocidal activity against tumor cells. Springerplus 4:569CrossRefGoogle Scholar
  24. Park B, Brinkmann MM, Spooner E, Lee CC, Kim YM, Ploegh HL (2008) Proteolytic cleavage in an endolysosomal compartment is required for activation of Toll-like receptor 9. Nat Immunol 9(12):1407–1414CrossRefGoogle Scholar
  25. Rawlings ND, Barrett AJ, Finn R (2016) Twenty years of the MEROPS database of proteolyticenzymes, their substrates and inhibitor. Nucleic Acids Res 44(D1):D343–D350CrossRefGoogle Scholar
  26. Reiser J, Adair B, Reinheckel T (2010) Specialized roles for cysteine cathepsins in health and disease. J Clin Invest 120(10):3421–3431CrossRefGoogle Scholar
  27. Riese RJ, Chapman HA (2000) Cathepsins and compartmentalization in antigen presentation. Curr Opin Immunol 12(1):107–113CrossRefGoogle Scholar
  28. Rossi A, Deveraux Q, Turk B, Sali A (2004) Comprehensive search for cysteine cathepsins in the human genome. Biol Chem 385:363–372CrossRefGoogle Scholar
  29. Smith JJ, Kuraku S, Holt C, Sauka-Spengler T, Li W et al (2013) Sequencing of the sea lamprey (Petromyzon marinus) genome provides insights into vertebrate evolution. Nat Genet 45(4):415–421 421e1-412CrossRefGoogle Scholar
  30. Stoka V, Turk V, Turk B (2016) Lysosomal cathepsins and their regulation in aging and neurodegeneration. Ageing Res Rev 32:22–37CrossRefGoogle Scholar
  31. Turk B, Turk D, Salvesen GS (2002) Regulating cysteine protease activity: essential role of protease inhibitors as guardians and regulators. Curr Pharm Des 8(18):1623–1637CrossRefGoogle Scholar
  32. Uinuk-Ool TS, Takezaki N, Kuroda N, Figueroa F, Sato A, Samonte IE, Mayer WE, Klein J (2003) Phylogeny of antigen-processing enzymes: cathepsins of a cephalochordate, an agnathan and a bony fish. Scand J Immunol 58(4):436–448CrossRefGoogle Scholar
  33. Unanue ER, Turk V, Neefjes J (2016) Variations in MHC class II antigen processing and presentation in health and disease. Annu Rev Immunol 34:265–297CrossRefGoogle Scholar
  34. Watts C (1997) Capture and processing of exogenous antigens for presentation on MHC molecules. Annu Rev Immunol 15:821–850CrossRefGoogle Scholar
  35. Willstätter R, Bamann E (1929) Über die Proteasen der Magenschleimhaut. Erste Abhandlung über die Enzyme der Leukocyten. Hoppe Seylers Z Physiol Chem 180:127–143CrossRefGoogle Scholar
  36. Xiao R, Zhang Z, Wang H, Han Y, Gou M, Li B, Duan D, Wang J, Liu X, Li Q (2015) Identification and characterization of a cathepsin D homologue from lampreys (Lampetra japonica). Dev Comp Immunol 49(1):149–156CrossRefGoogle Scholar
  37. Zavasnik-Bergant T, Turk B (2006) Cysteine cathepsins in the immune response. Tissue Antigens 67(5):349–355CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of Life ScienceLiaoning Normal UniversityDalianChina
  2. 2.Lamprey Research CenterLiaoning Normal UniversityDalianChina

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