MCA2_TRYB2
ID MCA2_TRYB2 Reviewed; 347 AA.
AC Q585F3; D6XHH8;
DT 07-OCT-2020, integrated into UniProtKB/Swiss-Prot.
DT 10-MAY-2005, sequence version 1.
DT 03-AUG-2022, entry version 99.
DE RecName: Full=Metacaspase-2 {ECO:0000305};
DE EC=3.4.22.- {ECO:0000269|PubMed:18005666, ECO:0000269|PubMed:22529389, ECO:0000269|PubMed:23506317, ECO:0000269|PubMed:28089596};
DE AltName: Full=TbMCA2 {ECO:0000303|PubMed:23506317};
DE Contains:
DE RecName: Full=Large subunit p20 {ECO:0000305};
DE Contains:
DE RecName: Full=Small subunit p10 {ECO:0000305};
DE Flags: Precursor;
GN Name=MCA2 {ECO:0000303|PubMed:18005666};
GN ORFNames=Tb927.6.940 {ECO:0000312|EMBL:AAX80349.1};
OS Trypanosoma brucei brucei (strain 927/4 GUTat10.1).
OC Eukaryota; Discoba; Euglenozoa; Kinetoplastea; Metakinetoplastina;
OC Trypanosomatida; Trypanosomatidae; Trypanosoma.
OX NCBI_TaxID=185431 {ECO:0000312|Proteomes:UP000008524};
RN [1] {ECO:0000312|EMBL:AAZ11657.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=927/4 GUTat10.1 {ECO:0000312|EMBL:AAZ11657.1};
RX PubMed=16020724; DOI=10.1126/science.1112181;
RA El-Sayed N.M., Myler P.J., Blandin G., Berriman M., Crabtree J.,
RA Aggarwal G., Caler E., Renauld H., Worthey E.A., Hertz-Fowler C.,
RA Ghedin E., Peacock C., Bartholomeu D.C., Haas B.J., Tran A.N.,
RA Wortman J.R., Alsmark U.C., Angiuoli S., Anupama A., Badger J.,
RA Bringaud F., Cadag E., Carlton J.M., Cerqueira G.C., Creasy T.,
RA Delcher A.L., Djikeng A., Embley T.M., Hauser C., Ivens A.C.,
RA Kummerfeld S.K., Pereira-Leal J.B., Nilsson D., Peterson J., Salzberg S.L.,
RA Shallom J., Silva J.C., Sundaram J., Westenberger S., White O.,
RA Melville S.E., Donelson J.E., Andersson B., Stuart K.D., Hall N.;
RT "Comparative genomics of trypanosomatid parasitic protozoa.";
RL Science 309:404-409(2005).
RN [2] {ECO:0000312|Proteomes:UP000008524}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=927/4 GUTat10.1 {ECO:0000312|Proteomes:UP000008524};
RX PubMed=16020726; DOI=10.1126/science.1112642;
RA Berriman M., Ghedin E., Hertz-Fowler C., Blandin G., Renauld H.,
RA Bartholomeu D.C., Lennard N.J., Caler E., Hamlin N.E., Haas B., Bohme U.,
RA Hannick L., Aslett M.A., Shallom J., Marcello L., Hou L., Wickstead B.,
RA Alsmark U.C.M., Arrowsmith C., Atkin R.J., Barron A.J., Bringaud F.,
RA Brooks K., Carrington M., Cherevach I., Chillingworth T.J., Churcher C.,
RA Clark L.N., Corton C.H., Cronin A., Davies R.M., Doggett J., Djikeng A.,
RA Feldblyum T., Field M.C., Fraser A., Goodhead I., Hance Z., Harper D.,
RA Harris B.R., Hauser H., Hostetler J., Ivens A., Jagels K., Johnson D.,
RA Johnson J., Jones K., Kerhornou A.X., Koo H., Larke N., Landfear S.,
RA Larkin C., Leech V., Line A., Lord A., Macleod A., Mooney P.J., Moule S.,
RA Martin D.M., Morgan G.W., Mungall K., Norbertczak H., Ormond D., Pai G.,
RA Peacock C.S., Peterson J., Quail M.A., Rabbinowitsch E., Rajandream M.A.,
RA Reitter C., Salzberg S.L., Sanders M., Schobel S., Sharp S., Simmonds M.,
RA Simpson A.J., Tallon L., Turner C.M., Tait A., Tivey A.R., Van Aken S.,
RA Walker D., Wanless D., Wang S., White B., White O., Whitehead S.,
RA Woodward J., Wortman J., Adams M.D., Embley T.M., Gull K., Ullu E.,
RA Barry J.D., Fairlamb A.H., Opperdoes F., Barrell B.G., Donelson J.E.,
RA Hall N., Fraser C.M., Melville S.E., El-Sayed N.M.A.;
RT "The genome of the African trypanosome Trypanosoma brucei.";
RL Science 309:416-422(2005).
RN [3] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, PROTEOLYTIC CLEAVAGE,
RP ACTIVE SITE, AND MUTAGENESIS OF LYS-55; CYS-212; CYS-213 AND LYS-268.
RX PubMed=18005666; DOI=10.1016/j.febslet.2007.11.009;
RA Moss C.X., Westrop G.D., Juliano L., Coombs G.H., Mottram J.C.;
RT "Metacaspase 2 of Trypanosoma brucei is a calcium-dependent cysteine
RT peptidase active without processing.";
RL FEBS Lett. 581:5635-5639(2007).
RN [4] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND DOMAIN.
RX PubMed=23506317; DOI=10.1111/febs.12248;
RA Machado M.F., Marcondes M.F., Juliano M.A., McLuskey K., Mottram J.C.,
RA Moss C.X., Juliano L., Oliveira V.;
RT "Substrate specificity and the effect of calcium on Trypanosoma brucei
RT metacaspase 2.";
RL FEBS J. 280:2608-2621(2013).
RN [5] {ECO:0000305}
RP CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES,
RP PROTEOLYTIC CLEAVAGE, AND MUTAGENESIS OF LYS-55 AND LYS-268.
RX PubMed=28089596; DOI=10.1016/j.bbapap.2017.01.002;
RA Gilio J.M., Marcondes M.F., Ferrari D., Juliano M.A., Juliano L.,
RA Oliveira V., Machado M.F.M.;
RT "Processing of metacaspase 2 from Trypanosoma brucei (TbMCA2) broadens its
RT substrate specificity.";
RL Biochim. Biophys. Acta 1865:388-394(2017).
RN [6] {ECO:0007744|PDB:4AF8, ECO:0007744|PDB:4AFP, ECO:0007744|PDB:4AFR}
RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) OF MUTANTS GLY-213 AND ALA-213,
RP CATALYTIC ACTIVITY, ACTIVITY REGULATION, SUBUNIT, PROTEOLYTIC CLEAVAGE, AND
RP MUTAGENESIS OF TYR-31; CYS-92; ASP-95; SER-156; 189-ASP--ASP-190 AND
RP ASP-211.
RX PubMed=22529389; DOI=10.1073/pnas.1200885109;
RA McLuskey K., Rudolf J., Proto W.R., Isaacs N.W., Coombs G.H., Moss C.X.,
RA Mottram J.C.;
RT "Crystal structure of a Trypanosoma brucei metacaspase.";
RL Proc. Natl. Acad. Sci. U.S.A. 109:7469-7474(2012).
CC -!- FUNCTION: Cysteine protease that cleaves specifically after arginine or
CC lysine residues. {ECO:0000269|PubMed:18005666,
CC ECO:0000269|PubMed:23506317}.
CC -!- ACTIVITY REGULATION: Activated by Ca(2+) (PubMed:18005666,
CC PubMed:23506317, PubMed:28089596, PubMed:22529389). In response to
CC calcium binding, the 280-loop, a disordered loop consisting of residues
CC 269-275, undergoes a conformational change which stabilizes substrates
CC in the active site (PubMed:22529389). The binding to the substrate
CC triggers the release of the N-terminal region resulting in the
CC activation of the enzyme (PubMed:22529389). Proteolytic cleavage is
CC required for catalytic activity towards large protein substrates
CC (PubMed:28089596). {ECO:0000269|PubMed:18005666,
CC ECO:0000269|PubMed:22529389, ECO:0000269|PubMed:23506317,
CC ECO:0000269|PubMed:28089596}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 7.7. {ECO:0000269|PubMed:23506317,
CC ECO:0000269|PubMed:28089596};
CC Temperature dependence:
CC Optimum temperature is 20-25 degrees Celsius.
CC {ECO:0000269|PubMed:23506317};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:22529389}.
CC -!- SUBCELLULAR LOCATION: Recycling endosome
CC {ECO:0000250|UniProtKB:Q8T8E7}.
CC -!- DOMAIN: There are 2 calcium binding sites with high and low affinity,
CC respectively. {ECO:0000269|PubMed:23506317}.
CC -!- PTM: Auto-proteolytic cleavage of the propeptide after Lys-55 and
CC between the large and small subunits after Lys-268 is required for
CC catalytic activity towards large protein substrates but is dispensable
CC towards small oligopeptide substrates (PubMed:18005666,
CC PubMed:28089596, PubMed:22529389). After processing, the propeptide and
CC the large and small subunits remain associated by non-covalent bonds
CC (PubMed:18005666). In vivo, the unprocessed enzyme appears to be the
CC predominant form (By similarity). {ECO:0000250|UniProtKB:Q8T8E7,
CC ECO:0000269|PubMed:18005666, ECO:0000269|PubMed:22529389,
CC ECO:0000269|PubMed:28089596}.
CC -!- SIMILARITY: Belongs to the peptidase C14B family. {ECO:0000305}.
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DR EMBL; CP000069; AAZ11657.1; -; Genomic_DNA.
DR EMBL; AC073906; AAX80349.1; -; Genomic_DNA.
DR RefSeq; XP_845216.1; XM_840123.1.
DR PDB; 4AF8; X-ray; 1.40 A; A=1-347.
DR PDB; 4AFP; X-ray; 2.10 A; A=1-347.
DR PDB; 4AFR; X-ray; 1.60 A; A=1-347.
DR PDB; 4AFV; X-ray; 1.50 A; A=1-347.
DR PDBsum; 4AF8; -.
DR PDBsum; 4AFP; -.
DR PDBsum; 4AFR; -.
DR PDBsum; 4AFV; -.
DR AlphaFoldDB; Q585F3; -.
DR SMR; Q585F3; -.
DR STRING; 5691.AAZ11657; -.
DR MEROPS; C14.044; -.
DR PaxDb; Q585F3; -.
DR GeneID; 3657732; -.
DR KEGG; tbr:Tb927.6.940; -.
DR VEuPathDB; TriTrypDB:Tb927.6.940; -.
DR eggNOG; KOG1546; Eukaryota.
DR InParanoid; Q585F3; -.
DR OMA; PTKANMI; -.
DR Proteomes; UP000008524; Chromosome 6.
DR GO; GO:0005634; C:nucleus; IDA:GeneDB.
DR GO; GO:0055037; C:recycling endosome; IEA:UniProtKB-SubCell.
DR GO; GO:0004197; F:cysteine-type endopeptidase activity; IDA:UniProtKB.
DR GO; GO:0008234; F:cysteine-type peptidase activity; IDA:GeneDB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0006508; P:proteolysis; IDA:UniProtKB.
DR InterPro; IPR029030; Caspase-like_dom_sf.
DR SUPFAM; SSF52129; SSF52129; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Autocatalytic cleavage; Calcium; Endosome; Hydrolase;
KW Metal-binding; Protease; Reference proteome; Thiol protease; Zymogen.
FT PROPEP 1..55
FT /evidence="ECO:0000269|PubMed:18005666"
FT /id="PRO_0000451273"
FT CHAIN 56..347
FT /note="Metacaspase-2"
FT /id="PRO_0000451274"
FT CHAIN 56..268
FT /note="Large subunit p20"
FT /evidence="ECO:0000269|PubMed:18005666"
FT /id="PRO_0000451275"
FT CHAIN 269..347
FT /note="Small subunit p10"
FT /evidence="ECO:0000269|PubMed:18005666"
FT /id="PRO_0000451276"
FT REGION 1..70
FT /note="Regulates substrate access to the active site"
FT /evidence="ECO:0000305|PubMed:22529389"
FT ACT_SITE 158
FT /evidence="ECO:0000250|UniProtKB:Q08601"
FT ACT_SITE 213
FT /evidence="ECO:0000305|PubMed:18005666"
FT BINDING 173
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:22529389,
FT ECO:0007744|PDB:4AFP"
FT BINDING 189
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:22529389,
FT ECO:0007744|PDB:4AFP"
FT BINDING 190
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:22529389,
FT ECO:0007744|PDB:4AFP"
FT BINDING 220
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:22529389,
FT ECO:0007744|PDB:4AFP"
FT SITE 55..56
FT /note="Cleavage; by autolysis"
FT /evidence="ECO:0000269|PubMed:18005666"
FT SITE 95
FT /note="Important for Arg/Lys-specific substrate
FT specificity"
FT /evidence="ECO:0000269|PubMed:22529389"
FT SITE 211
FT /note="Important for Arg/Lys-specific substrate
FT specificity"
FT /evidence="ECO:0000269|PubMed:22529389"
FT SITE 268
FT /note="Cleavage; by autolysis"
FT /evidence="ECO:0000269|PubMed:18005666"
FT MUTAGEN 31
FT /note="Y->A: Increases autoprocessing resulting in the
FT degradation of the enzyme."
FT /evidence="ECO:0000269|PubMed:22529389"
FT MUTAGEN 55
FT /note="K->G: Loss of autoprocessing. Loss of catalytic
FT activity towards large protein substrates, no effect on
FT catalytic activity towards short oligopeptide substrates,
FT reduces affinity of the high affinity Ca(2+) binding site,
FT shifts optimum pH for activity towards basic pH; when
FT associated with G-268."
FT /evidence="ECO:0000269|PubMed:18005666,
FT ECO:0000269|PubMed:28089596"
FT MUTAGEN 92
FT /note="C->A: Reduced autoprocessing and 50% loss of
FT catalytic activity towards substrates."
FT /evidence="ECO:0000269|PubMed:22529389"
FT MUTAGEN 95
FT /note="D->A: Loss of autoprocessing and catalytic activity
FT towards substrates."
FT /evidence="ECO:0000269|PubMed:22529389"
FT MUTAGEN 156
FT /note="S->A: 3-fold increase in catalytic activity towards
FT substrates."
FT /evidence="ECO:0000269|PubMed:22529389"
FT MUTAGEN 189..190
FT /note="DD->AA: Loss of autoprocessing and catalytic
FT activity towards substrates."
FT /evidence="ECO:0000269|PubMed:22529389"
FT MUTAGEN 211
FT /note="D->A: Loss of autoprocessing and catalytic activity
FT towards substrates."
FT /evidence="ECO:0000269|PubMed:22529389"
FT MUTAGEN 212
FT /note="C->G: Severe loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:18005666"
FT MUTAGEN 213
FT /note="C->G: Complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:18005666"
FT MUTAGEN 268
FT /note="K->G: Loss of autoprocessing. Loss of catalytic
FT activity towards large protein substrates, no effect on
FT catalytic activity towards short oligopeptide substrates,
FT reduces affinity of the high affinity Ca(2+) binding site,
FT shift optimum pH for activity towards basic pH; when
FT associated with G-55."
FT /evidence="ECO:0000269|PubMed:18005666,
FT ECO:0000269|PubMed:28089596"
FT TURN 8..10
FT /evidence="ECO:0007829|PDB:4AFR"
FT TURN 17..24
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 31..34
FT /evidence="ECO:0007829|PDB:4AFV"
FT HELIX 47..54
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 73..79
FT /evidence="ECO:0007829|PDB:4AF8"
FT HELIX 92..105
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 111..117
FT /evidence="ECO:0007829|PDB:4AF8"
FT HELIX 131..143
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 150..157
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 159..162
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 173..176
FT /evidence="ECO:0007829|PDB:4AF8"
FT HELIX 181..184
FT /evidence="ECO:0007829|PDB:4AF8"
FT HELIX 189..196
FT /evidence="ECO:0007829|PDB:4AF8"
FT TURN 197..199
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 205..211
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 222..226
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 250..258
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 278..281
FT /evidence="ECO:0007829|PDB:4AFV"
FT HELIX 283..293
FT /evidence="ECO:0007829|PDB:4AF8"
FT HELIX 300..313
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 319..326
FT /evidence="ECO:0007829|PDB:4AF8"
FT STRAND 336..338
FT /evidence="ECO:0007829|PDB:4AF8"
SQ SEQUENCE 347 AA; 37779 MW; 00523E8CCF7B214A CRC64;
MCSLITQLCD AGQLADYVGL GWLNAVSSQP YLVQALGLQP PPRRVDVDAA FRDAKGLHGH
QPWVATPLPG QTVRALFIGI NYYGTSAALS GCCNDVKQML ATLQKKGLPI NEAVILVDED
NFPGRTDQPT RDNIVRYMAW LVKDAKPGDV LFFHYSGHGT QCKSRGDSDE KYDQCIAPVD
FQKSGCIVDD DIHKLLFSRL PEKVRLTAVF DCCHSGSIMD LPFTYVCSGG EQASGTPHMK
RIREGNDVLG DVMMISGCAD EQTSADVKNT ATFGTGSTGA GGAATQCITC MLMNNQSLSY
GKLLIETRDM LKRKGFKQVP QLSASKAIDL DQTFSLTEMF SVDRSIQ
