Summary: A family of SERINE ENDOPEPTIDASES isolated from Bacillus subtilis. EC 3.4.21.-

Top Publications

  1. Vévodová J, Gamble M, Künze G, Ariza A, Dodson E, Jones D, et al. Crystal structure of an intracellular subtilisin reveals novel structural features unique to this subtilisin family. Structure. 2010;18:744-55 pubmed publisher
    ..The first ISP structure reported herein at a spacing of 1.56 A reveals features unique among subtilisins that has enabled potential functional and physiological roles to be assigned to sequence elements exclusive to ..
  2. Sanchez S, Díaz Sánchez S, Martínez R, Llorente M, Herrera Leon S, Vidal D. The new allelic variant of the subtilase cytotoxin (subAB2) is common among Shiga toxin-producing Escherichia coli strains from large game animals and their meat and meat products. Vet Microbiol. 2013;166:645-9 pubmed publisher
  3. Eppler R, Komor R, Huynh J, Dordick J, Reimer J, Clark D. Water dynamics and salt-activation of enzymes in organic media: mechanistic implications revealed by NMR spectroscopy. Proc Natl Acad Sci U S A. 2006;103:5706-10 pubmed
    ..This increased mobility may contribute to the dramatic increases in biocatalyst activity. ..
  4. Wang C, Typas M, Butt T. Phylogenetic and exon-intron structure analysis of fungal subtilisins: support for a mixed model of intron evolution. J Mol Evol. 2005;60:238-46 pubmed
    Phylogenetic and exon-intron structure analyses of intra- and interspecific fungal subtilisins in this study provided support for a mixed model of intron evolution: a synthetic theory of introns-early and introns-late speculations...
  5. Byres E, Paton A, Paton J, L fling J, Smith D, Wilce M, et al. Incorporation of a non-human glycan mediates human susceptibility to a bacterial toxin. Nature. 2008;456:648-52 pubmed publisher
    ..Ironically, foods rich in Neu5Gc are the most common source of STEC contamination. Thus a bacterial toxin's receptor is generated by metabolic incorporation of an exogenous factor derived from food...
  6. Takeda N, Sato S, Asamizu E, Tabata S, Parniske M. Apoplastic plant subtilases support arbuscular mycorrhiza development in Lotus japonicus. Plant J. 2009;58:766-77 pubmed publisher
    ..Our data indicate a role for these subtilases during the fungal infection process in particular arbuscule development. ..
  7. Morinaga N, Yahiro K, Matsuura G, Moss J, Noda M. Subtilase cytotoxin, produced by Shiga-toxigenic Escherichia coli, transiently inhibits protein synthesis of Vero cells via degradation of BiP and induces cell cycle arrest at G1 by downregulation of cyclin D1. Cell Microbiol. 2008;10:921-9 pubmed
    ..SubAB-treated cells showed cell cycle arrest in G1 phase, which may result from cyclin D1 downregulation caused by both SubAB-induced translational inhibition and continuous prolonged proteasomal degradation. ..
  8. Bielaszewska M, Karch H. Consequences of enterohaemorrhagic Escherichia coli infection for the vascular endothelium. Thromb Haemost. 2005;94:312-8 pubmed
    ..Based on current data, cytolethal distending toxin, EHEC haemolysin, and subtilase cytotoxin might be such candidates...
  9. May K, Paton J, Paton A. Escherichia coli subtilase cytotoxin induces apoptosis regulated by host Bcl-2 family proteins Bax/Bak. Infect Immun. 2010;78:4691-6 pubmed publisher

More Information


  1. Koussis K, Withers Martinez C, Yeoh S, Child M, Hackett F, Knuepfer E, et al. A multifunctional serine protease primes the malaria parasite for red blood cell invasion. EMBO J. 2009;28:725-35 pubmed publisher
    ..We propose that PfSUB1 is a multifunctional processing protease with an essential role in both egress of the malaria merozoite and remodelling of its surface in preparation for erythrocyte invasion. ..
  2. Baldo A, Tabart J, Vermout S, Mathy A, Collard A, Losson B, et al. Secreted subtilisins of Microsporum canis are involved in adherence of arthroconidia to feline corneocytes. J Med Microbiol. 2008;57:1152-6 pubmed publisher
    ..Moreover, two mAbs against the keratinolytic protease subtilisin 3 (Sub3) inhibited M. canis adherence to RFE by 23%, suggesting that subtilisins, and Sub3 in particular, are involved in the adherence process.
  3. Jones D. Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. PLoS ONE. 2011;6:e24319 pubmed publisher
    ..fragment exchange was utilised to recombine key structural and functional low homology regions of bacilli subtilisins to generate new active hybrid proteases with altered substrate profiles...
  4. Ganesan A, Price N, Kelly S, Petry I, Moore B, Halling P. Circular dichroism studies of subtilisin Carlsberg immobilised on micron sized silica particles. Biochim Biophys Acta. 2006;1764:1119-25 pubmed
    ..Additional evidence for loss of native conformation is provided by the significant decrease in the near UV CD spectrum. These results for the first time clearly demonstrate the origin of enzyme instability in the immobilised state. ..
  5. Ruan B, Fisher K, Alexander P, Doroshko V, Bryan P. Engineering subtilisin into a fluoride-triggered processing protease useful for one-step protein purification. Biochemistry. 2004;43:14539-46 pubmed
    ..Cleavage can be triggered by the addition of fluoride to release the pure target protein. The column is then regenerated by stripping off the tightly bound prodomain at pH 2.1. Ten proteins have been purified to date by this method. ..
  6. Matsuura G, Morinaga N, Yahiro K, Komine R, Moss J, Yoshida H, et al. Novel subtilase cytotoxin produced by Shiga-toxigenic Escherichia coli induces apoptosis in vero cells via mitochondrial membrane damage. Infect Immun. 2009;77:2919-24 pubmed publisher
    ..These data suggested that SubAB induced caspase-dependent apoptosis in Vero cells through mitochondrial membrane damage. ..
  7. Withers Martinez C, Jean L, Blackman M. Subtilisin-like proteases of the malaria parasite. Mol Microbiol. 2004;53:55-63 pubmed
    ..They may have potential as novel drug targets. Here, we review progress in our understanding of the maturation, specificity, structure and function of these Plasmodium subtilases. ..
  8. Withers Martinez C, Suarez C, Fulle S, Kher S, Penzo M, Ebejer J, et al. Plasmodium subtilisin-like protease 1 (SUB1): insights into the active-site structure, specificity and function of a pan-malaria drug target. Int J Parasitol. 2012;42:597-612 pubmed publisher
    ..Our findings demonstrate that it should be possible to develop 'pan-reactive' drug-like compounds that inhibit SUB1 in all three major human malaria pathogens, enabling production of broad-spectrum antimalarial drugs targeting SUB1...
  9. Wolfson J, May K, Thorpe C, Jandhyala D, Paton J, Paton A. Subtilase cytotoxin activates PERK, IRE1 and ATF6 endoplasmic reticulum stress-signalling pathways. Cell Microbiol. 2008;10:1775-86 pubmed publisher
    ..The rapidity with which ER stress-signalling responses are triggered by exposure of cells to SubAB is consistent with the hypothesis that cleavage by the toxin causes BiP to dissociate from the signalling molecules. ..
  10. Georgieva D, Genov N, Betzel C. Bacillus licheniformis variant DY proteinase: specificity in relation to the geometry of the substrate recognition site. Curr Microbiol. 2005;51:71-4 pubmed
    ..The results characterize subtilisin DY as a bacterial proteinase with a broad specificity due to the specific geometry and flexibility of the substrate recognition site, which can accommodate different types of amino acid side chains. ..
  11. Hu G, Leger R. A phylogenomic approach to reconstructing the diversification of serine proteases in fungi. J Evol Biol. 2004;17:1204-14 pubmed
    ..A similar prevailing trend towards lineage specific gene loss of trypsins in saprophytes and some pathogens suggests that their phylogenetic breadth will have been much wider in early fungi than currently. ..
  12. Radisky E, Kwan G, Karen Lu C, Koshland D. Binding, proteolytic, and crystallographic analyses of mutations at the protease-inhibitor interface of the subtilisin BPN'/chymotrypsin inhibitor 2 complex. Biochemistry. 2004;43:13648-56 pubmed
  13. Cedzich A, Huttenlocher F, Kuhn B, Pfannstiel J, Gabler L, Stintzi A, et al. The protease-associated domain and C-terminal extension are required for zymogen processing, sorting within the secretory pathway, and activity of tomato subtilase 3 (SlSBT3). J Biol Chem. 2009;284:14068-78 pubmed publisher
    ..Both features were found to be required for SlSBT3 activity and, consequently, for prodomain processing and secretion...
  14. Wang H, Paton J, Thorpe C, Bonder C, Sun W, Paton A. Tissue factor–dependent procoagulant activity of subtilase cytotoxin, a potent AB5 toxin produced by shiga toxigenic Escherichia coli. J Infect Dis. 2010;202:1415-23 pubmed publisher
    ..Our results suggest that the procoagulant effect of SubAB may be dependent on both the up?regulation of TF expression and the activation of TF by means of BiP cleavage. ..
  15. Gerhardt E, Masso M, Paton A, Paton J, Zotta E, Ibarra C. Inhibition of water absorption and selective damage to human colonic mucosa are induced by subtilase cytotoxin produced by Escherichia coli O113:H21. Infect Immun. 2013;81:2931-7 pubmed publisher
    ..coli O113:H21 expressing only SubAB was assayed. This is the first study showing that SubAB may directly participate in the mechanisms of diarrhea in children infected with non-O157 STEC strains. ..
  16. Maurer K. Detergent proteases. Curr Opin Biotechnol. 2004;15:330-4 pubmed
    Over the past 20 years, the development of subtilisins as typical detergent proteases has employed all the tools of enzyme technology, resulting in a constant flow of new and improved enzymes...
  17. Michelacci V, Tozzoli R, Caprioli A, Martinez R, Scheutz F, Grande L, et al. A new pathogenicity island carrying an allelic variant of the Subtilase cytotoxin is common among Shiga toxin producing Escherichia coli of human and ovine origin. Clin Microbiol Infect. 2013;19:E149-56 pubmed publisher
    ..Further studies are needed to assess the role of tia and/or other genes carried by SE-PAI in the colonization of the host intestinal mucosa. ..
  18. Bryant M, Schardl C, Hesse U, Scott B. Evolution of a subtilisin-like protease gene family in the grass endophytic fungus Epichloë festucae. BMC Evol Biol. 2009;9:168 pubmed publisher
    ..Phylogenetic analysis in Hypocreales fungi revealed an extensive history of gene loss and duplication. This study provides new insights into the evolution of the SLP superfamily in filamentous fungi. ..
  19. Xue A, Chapoval S, Finn E, Chowdhary V, Marietta E, Gaffey T, et al. HLA-DQ8 is a predisposing molecule for detergent enzyme subtilisin BPN'-induced hypersensitivity. Clin Immunol. 2005;117:302-15 pubmed
  20. Yeoh S, O Donnell R, Koussis K, Dluzewski A, Ansell K, Osborne S, et al. Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Cell. 2007;131:1072-83 pubmed
    ..Our findings reveal the presence in the malarial parasitophorous vacuole of a regulated, PfSUB1-mediated proteolytic processing event required for release of viable parasites from the host erythrocyte. ..
  21. Lass A, Kujawa M, McConnell E, Paton A, Paton J, Wojcik C. Decreased ER-associated degradation of alpha-TCR induced by Grp78 depletion with the SubAB cytotoxin. Int J Biochem Cell Biol. 2008;40:2865-79 pubmed publisher
    ..Expression of CHOP/GADD153 occurred only after prolonged incubation and was not associated with apoptosis. ..
  22. Arastu Kapur S, Ponder E, Fonović U, Yeoh S, Yuan F, Fonovic M, et al. Identification of proteases that regulate erythrocyte rupture by the malaria parasite Plasmodium falciparum. Nat Chem Biol. 2008;4:203-13 pubmed publisher
    ..These results suggest that two mechanistically distinct proteases function to regulate processing of downstream substrates required for efficient release of parasites from host red blood cells. ..
  23. Pereira M, Byrne B, Nguyen T, Lewis D, Atwill E. The occurrence of subtilase-cytotoxin-encoding genes in environmental Escherichia coli isolated from a Northern California estuary. Can J Microbiol. 2013;59:437-41 pubmed publisher
    ..To further understand potential health risks posed by strains encoding SubAB, future epidemiological studies should consider screening isolates for subAB regardless of the presence of Shiga-toxin-encoding genes. ..
  24. Sanchez S, Beristain X, Martínez R, Garcia A, Martin C, Vidal D, et al. Subtilase cytotoxin encoding genes are present in human, sheep and deer intimin-negative, Shiga toxin-producing Escherichia coli O128:H2. Vet Microbiol. 2012;159:531-5 pubmed publisher
    ..Our results indicate that sheep and deer represent a reservoir of SubAB-positive STEC O128:H2 strains and thus a potential source of human infection. ..
  25. Rautengarten C, Steinhauser D, Büssis D, Stintzi A, Schaller A, Kopka J, et al. Inferring hypotheses on functional relationships of genes: Analysis of the Arabidopsis thaliana subtilase gene family. PLoS Comput Biol. 2005;1:e40 pubmed
    ..Supplemental material is available in the Plant Subtilase Database (PSDB) (, as well as from the CSB.DB ( ..
  26. Shinde U, Thomas G. Insights from bacterial subtilases into the mechanisms of intramolecular chaperone-mediated activation of furin. Methods Mol Biol. 2011;768:59-106 pubmed publisher
    Prokaryotic subtilisins and eukaryotic proprotein convertases (PCs) are two homologous protease subfamilies that belong to the larger ubiquitous super-family called subtilases...
  27. Schaller A, Stintzi A, Graff L. Subtilases - versatile tools for protein turnover, plant development, and interactions with the environment. Physiol Plant. 2012;145:52-66 pubmed publisher
    ..In this article we provide an overview of the earlier literature on the discovery of the first SBTs in plants, and highlight recent findings with respect to their physiological relevance, structure and function. ..
  28. Faraco V, Palmieri G, Festa G, Monti M, Sannia G, Giardina P. A new subfamily of fungal subtilases: structural and functional analysis of a Pleurotus ostreatus member. Microbiology. 2005;151:457-66 pubmed
    ..A new subgroup of subtilisin-like proteases, belonging to the pyrolysin family, has been defined, which includes proteases from ascomycete and basidiomycete fungi. ..
  29. Chong D, Paton J, Thorpe C, Paton A. Clathrin-dependent trafficking of subtilase cytotoxin, a novel AB5 toxin that targets the endoplasmic reticulum chaperone BiP. Cell Microbiol. 2008;10:795-806 pubmed
  30. Tsutsuki H, Yahiro K, Suzuki K, Suto A, Ogura K, Nagasawa S, et al. Subtilase cytotoxin enhances Escherichia coli survival in macrophages by suppression of nitric oxide production through the inhibition of NF-?B activation. Infect Immun. 2012;80:3939-51 pubmed publisher
    ..coli survival in macrophages. Thus, we hypothesize that SubAB might represent a novel bacterial strategy to circumvent host defense during STEC infection. ..
  31. Bonifait L, Vaillancourt K, Gottschalk M, Frenette M, Grenier D. Purification and characterization of the subtilisin-like protease of Streptococcus suis that contributes to its virulence. Vet Microbiol. 2011;148:333-40 pubmed publisher
    ..In conclusion, the SspA of S. suis shared similarities with subtilisin-like proteases produced by other pathogenic streptococci and may contribute to the pathogenic process of S. suis infections. ..
  32. Yahiro K, Morinaga N, Moss J, Noda M. Subtilase cytotoxin induces apoptosis in HeLa cells by mitochondrial permeabilization via activation of Bax/Bak, independent of C/EBF-homologue protein (CHOP), Ire1alpha or JNK signaling. Microb Pathog. 2010;49:153-63 pubmed publisher
  33. Tozzoli R, Caprioli A, Cappannella S, Michelacci V, Marziano M, Morabito S. Production of the subtilase AB5 cytotoxin by Shiga toxin-negative Escherichia coli. J Clin Microbiol. 2010;48:178-83 pubmed publisher
    ..These results indicate that SubAB production is not restricted to STEC and suggest that a subAB-tia putative pathogenicity island is involved in the dissemination of subAB genes, as an alternative to plasmid pO113. ..
  34. Löfling J, Paton A, Varki N, Paton J, Varki A. A dietary non-human sialic acid may facilitate hemolytic-uremic syndrome. Kidney Int. 2009;76:140-4 pubmed publisher
  35. Jean F, Thomas L, Molloy S, Liu G, Jarvis M, Nelson J, et al. A protein-based therapeutic for human cytomegalovirus infection. Proc Natl Acad Sci U S A. 2000;97:2864-9 pubmed
  36. Bagga S, Hu G, Screen S, St Leger R. Reconstructing the diversification of subtilisins in the pathogenic fungus Metarhizium anisopliae. Gene. 2004;324:159-69 pubmed
    ..Here we sought a global characterization of the diversity of subtilisins in the insect pathogen Metarhizium anisopliae...
  37. Thomas G. Furin at the cutting edge: from protein traffic to embryogenesis and disease. Nat Rev Mol Cell Biol. 2002;3:753-66 pubmed
    ..This review summarizes various features of furin--its structural and enzymatic properties, intracellular localization, trafficking, substrates, and roles in vivo. ..
  38. Seidah N, Chretien M. Proprotein and prohormone convertases: a family of subtilases generating diverse bioactive polypeptides. Brain Res. 1999;848:45-62 pubmed
  39. Basak S, Chretien M, Mbikay M, Basak A. In vitro elucidation of substrate specificity and bioassay of proprotein convertase 4 using intramolecularly quenched fluorogenic peptides. Biochem J. 2004;380:505-14 pubmed
    ..It was also revealed that PC4 is a good candidate processing enzyme for growth factors IGF-1 and -2, neuropeptide proPACAP and several ADAM proteins such as ADAM-1, -2, -3 and -5. ..
  40. Rockwell N, Krysan D, Komiyama T, Fuller R. Precursor processing by kex2/furin proteases. Chem Rev. 2002;102:4525-48 pubmed
  41. Venancio E, Daher B, Andrade R, Soares C, Pereira I, Felipe M. The kex2 gene from the dimorphic and human pathogenic fungus Paracoccidioides brasiliensis. Yeast. 2002;19:1221-31 pubmed
    ..A putative Golgi retrieval signal (YEFEMI) has also been found in the cytoplasmic tail. The complete nucleotide sequence of Pbkex2 and its flanking regions have been submitted to GenBank database under Accession No. AF486805. ..
  42. Withers Martinez C, Saldanha J, Ely B, Hackett F, O Connor T, Blackman M. Expression of recombinant Plasmodium falciparum subtilisin-like protease-1 in insect cells. Characterization, comparison with the parasite protease, and homology modeling. J Biol Chem. 2002;277:29698-709 pubmed
    ..homology modeling of the PfSUB-1 catalytic domain based on an alignment with closely related bacterial subtilisins and an orthologue from the rodent malaria Plasmodium yoelii suggests that the protease has at least three ..
  43. Toogood H, Smith C, Baker E, Daniel R. Purification and characterization of Ak.1 protease, a thermostable subtilisin with a disulphide bond in the substrate-binding cleft. Biochem J. 2000;350 Pt 1:321-8 pubmed
    ..However, it was found that this protease has a limited substrate specificity, so this application was not explored further. ..
  44. Hamilton J, Simpson D, Hyman S, Ndimba B, Slabas A. Ara12 subtilisin-like protease from Arabidopsis thaliana: purification, substrate specificity and tissue localization. Biochem J. 2003;370:57-67 pubmed
    ..Possible functions of Ara12 are discussed in the light of the involvement of a number of plant subtilisin-like proteases in morphogenesis. ..
  45. Barrette Ng I, Ng K, Cherney M, Pearce G, Ryan C, James M. Structural basis of inhibition revealed by a 1:2 complex of the two-headed tomato inhibitor-II and subtilisin Carlsberg. J Biol Chem. 2003;278:24062-71 pubmed publisher
    ..subtilisin)2 complex provides a molecular framework for understanding how multiple inhibitory domains in a single Potato II type proteinase inhibitor molecule from the Potato II family act to inhibit proteolytic enzymes...
  46. Almog O, Gallagher D, Ladner J, Strausberg S, Alexander P, Bryan P, et al. Structural basis of thermostability. Analysis of stabilizing mutations in subtilisin BPN'. J Biol Chem. 2002;277:27553-8 pubmed
    ..The effects are attributed to the new disulfide cross-link and to improved hydrophobic packing, new hydrogen bonds, and other rearrangements in the N-terminal region. ..
  47. Jean L, Hackett F, Martin S, Blackman M. Functional characterization of the propeptide of Plasmodium falciparum subtilisin-like protease-1. J Biol Chem. 2003;278:28572-9 pubmed
    ..significant secondary structure on its own, comparable with that of folded propeptides of some bacterial subtilisins. Kinetic studies demonstrated that rp31 is a fast binding, high affinity inhibitor of PfSUB-1...
  48. Tanaka H, Onouchi H, Kondo M, Hara Nishimura I, Nishimura M, Machida C, et al. A subtilisin-like serine protease is required for epidermal surface formation in Arabidopsis embryos and juvenile plants. Development. 2001;128:4681-9 pubmed
  49. Takahashi M, Hasuura Y, Nakamori S, Takagi H. Improved autoprocessing efficiency of mutant subtilisins E with altered specificity by engineering of the pro-region. J Biochem. 2001;130:99-106 pubmed
    ..Therefore, to enhance processing, we engineered the pro-region of mutant subtilisins E of Bacillus subtilis with altered substrate specificity...
  50. Anderson E, Molloy S, Jean F, Fei H, Shimamura S, Thomas G. The ordered and compartment-specfific autoproteolytic removal of the furin intramolecular chaperone is required for enzyme activation. J Biol Chem. 2002;277:12879-90 pubmed
  51. Evans K, Crowder J, Miller E. Subtilisins of Bacillus spp. hydrolyze keratin and allow growth on feathers. Can J Microbiol. 2000;46:1004-11 pubmed
    ..PWD-1 and the kerA protease efficiently degrade keratin, keratin hydrolysis and growth on feathers is a property that can be conferred by appropriate expression of the major subtilisins, including the industrially produced enzymes.
  52. Uzureau P, Barale J, Janse C, Waters A, Breton C. Gene targeting demonstrates that the Plasmodium berghei subtilisin PbSUB2 is essential for red cell invasion and reveals spontaneous genetic recombination events. Cell Microbiol. 2004;6:65-78 pubmed
    ..berghei. ..
  53. Chen C, Huff M, Matteson J, Page L, Phillips R, Kelly J, et al. Furin initiates gelsolin familial amyloidosis in the Golgi through a defect in Ca(2+) stabilization. EMBO J. 2001;20:6277-87 pubmed
    ..We propose that coincidence of membrane trafficking pathways contributes to the development of proteolysis-initiated amyloid disease. ..
  54. Takagi H, Koga M, Katsurada S, Yabuta Y, Shinde U, Inouye M, et al. Functional analysis of the propeptides of subtilisin E and aqualysin I as intramolecular chaperones. FEBS Lett. 2001;508:210-4 pubmed
    ..Further, some enzymatic properties of some chimeras in which the subtilisin mature domain is partly swapped with that of aqualysin I were shown to be more similar to those of aqualysin I. ..
  55. Boyd P, Barnaby N, Tan Wilson A, Wilson K. Cleavage specificity of the subtilisin-like protease C1 from soybean. Biochim Biophys Acta. 2002;1596:269-82 pubmed
  56. Blackman M, Corrie J, Croney J, Kelly G, Eccleston J, Jameson D. Structural and biochemical characterization of a fluorogenic rhodamine-labeled malarial protease substrate. Biochemistry. 2002;41:12244-52 pubmed
    ..These data also indicate that the rhodamine dimers fluoresce and that the associated lifetimes are subnanosecond. ..