Reviewed, UniProtKB/Swiss-Prot P25779 (CYSP_TRYCR)

Names and origin

Protein names	Recommended name:     Cruzipain     EC=3.4.22.51 Alternative name(s):     Cruzaine     Major cysteine proteinase
Organism	Trypanosoma cruzi
Taxonomic identifier	5693 [NCBI]
Taxonomic lineage	Eukaryota › Euglenozoa › Kinetoplastida › Trypanosomatidae › Trypanosoma › Schizotrypanum

Protein attributes

Sequence length	467 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Hydrolyzes chromogenic peptides at the carboxyl Arg or Lys; requires at least one more amino acid, preferably Arg, Phe, Val or Leu, between the terminal Arg or Lys and the amino-blocking group. The cysteine protease may play an important role in the development and differentiation of the parasites at several stages of their life cycle.
Catalytic activity	Broad endopeptidase specificity similar to that of cathepsin L.
Enzyme regulation	Strongly inhibited by E-64 (L-trans-epoxysuccinylleucylamido(4-guanidino)butane), Leupeptin, and N-alpha-p-tosyl-L-lysine chloromethyl ketone.
Developmental stage	Present in all developmental stages.
Miscellaneous	Purified cruzipain is able to degrade itself, yielding a complex mixture of small peptides, and a major 25 kDa fragment.
Sequence similarities	Belongs to the peptidase C1 family.

Ontologies

Keywords
Domain	Signal
Molecular function	Hydrolase Protease Thiol protease
PTM	Autocatalytic cleavage Disulfide bond Glycoprotein Zymogen
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological process	proteolysis Inferred from electronic annotation. Source: InterPro
Molecular function	cysteine-type endopeptidase activity Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 18

18

Probable

Propeptide

19 – 122

104

Activation peptide Probable

PRO_0000026372

Chain

123 – 467

345

Cruzipain

PRO_0000026373

Sites

Active site

147

1

Active site

284

1

Active site

304

1

Site

337 – 338

2

Cleavage; by autolysis Ref.1

Amino acid modifications

Glycosylation

169

1

N-linked (GlcNAc...) Potential

Glycosylation

292

1

N-linked (GlcNAc...) Potential

Glycosylation

377

1

N-linked (GlcNAc...) Potential

Disulfide bond

144 ↔ 185

Disulfide bond

178 ↔ 223

Disulfide bond

277 ↔ 325

Natural variations

Natural variant

9

1

L → S in clone 1800-2.

Natural variant

35

1

T → A in clone 1800-2.

Natural variant

39

1

A → V in clone 1800-2.

Natural variant

51

1

S → N in clone 1800-4.

Natural variant

56

1

A → R in clone 1800-2.

Natural variant

76

1

N → G in clone 1800-4.

Natural variant

109

1

Q → G in clone 1800-4.

Natural variant

117

1

K → N in clone 1800-2.

Natural variant

146

1

S → G

Natural variant

157 – 158

2

EC → SG in strain: RA.

Natural variant

186

1

S → G in clones 1800-2 and 1800-4.

Natural variant

204

1

A → G in strain: RA.

Natural variant

250 – 251

2

WL → CV in clones 1800-2 and 1800-4.

Natural variant

261 – 262

2

VD → H in strain: RA.

Natural variant

286

1

V → F in clone 1800-4.

Natural variant

286

1

V → L in strain: RA.

Natural variant

308

1

T → A in clone 1800-2.

Natural variant

313

1

E → D

Natural variant

409

1

L → F in clone 1800-2.

Natural variant

427

1

K → Q in clone 1800-2.

Natural variant

430

1

R → W in clone 1800-2.

Natural variant

457

1

H → Y in clone 1800-2.

Natural variant

461

1

H → Q in clone 1800-2.

Experimental info

Sequence conflict

357

1

S → C Ref.4

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

467

Helix Strand Turn

Details...

Beta strand

125 – 128

4

Helix

129 – 132

4

Helix

147 – 162

16

Helix

172 – 178

7

Helix

190 – 200

11

Beta strand

204 – 207

4

Turn

208 – 210

3

Beta strand

230 – 233

4

Beta strand

236 – 239

4

Helix

244 – 254

11

Beta strand

257 – 261

5

Helix

266 – 268

3

Beta strand

271 – 274

4

Beta strand

284 – 297

14

Beta strand

299 – 303

5

Beta strand

315 – 322

8

Helix

324 – 326

3

Beta strand

329 – 335

7

Sequences

	Sequence		Length	Mass (Da)
	P25779-1 [UniParc]. Last modified May 1, 1992. Version 1. Checksum: B93EBA49B511363D	FASTA	467	49,836
	10 20 30 40 50 60 MSGWARALLL AAVLVVMACL VPAATASLHA EETLTSQFAE FKQKHGRVYE SAAEEAFRLS 70 80 90 100 110 120 VFRENLFLAR LHAAANPHAT FGVTPFSDLT REEFRSRYHN GAAHFAAAQE RARVPVKVEV 130 140 150 160 170 180 VGAPAAVDWR ARGAVTAVKD QGQCGSCWAF SAIGNVECQW FLAGHPLTNL SEQMLVSCDK 190 200 210 220 230 240 TDSGCSGGLM NNAFEWIVQE NNGAVYTEDS YPYASGEGIS PPCTTSGHTV GATITGHVEL 250 260 270 280 290 300 PQDEAQIAAW LAVNGPVAVA VDASSWMTYT GGVMTSCVSE QLDHGVLLVG YNDSAAVPYW 310 320 330 340 350 360 IIKNSWTTQW GEEGYIRIAK GSNQCLVKEE ASSAVVGGPG PTPEPTTTTT TSAPGPSPSY 370 380 390 400 410 420 FVQMSCTDAA CIVGCENVTL PTGQCLLTTS GVSAIVTCGA ETLTEEVFLT STHCSGPSVR 430 440 450 460 SSVPLNKCNR LLRGSVEFFC GSSSSGRLAD VDRQRRHQPY HSRHRRL

References

[1]	"The sequence, organization, and expression of the major cysteine protease (cruzain) from Trypanosoma cruzi." Eakin A.E., Mills A.A., Harth G., McKerrow J.H., Craik C.S. J. Biol. Chem. 267:7411-7420(1992) [PubMed: 1559982] [Abstract] Cited for: NUCLEOTIDE SEQUENCE. Strain: Tulahuen.
[2]	"The major cysteine proteinase (cruzipain) from Trypanosoma cruzi is encoded by multiple polymorphic tandemly organized genes located on different chromosomes." Campetella O., Henriksson J., Aaslund L., Frasch A.C.C., Pettersson U., Cazzulo J.J. Mol. Biochem. Parasitol. 50:225-234(1992) [PubMed: 1311053] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] (CLONES 1800-2 AND 1800-4). Strain: Tulahuen 2.
[3]	"Amplification and sequencing of genomic DNA fragments encoding cysteine proteases from protozoan parasites." Eakin A.E., Bouvier J., Sakanari J.A., Craik C.S., McKerrow J.H. Mol. Biochem. Parasitol. 39:1-8(1990) [PubMed: 2406590] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 141-306. Strain: RA.
[4]	"The C-terminal extension of the major cysteine proteinase (cruzipain) from Trypanosoma cruzi." Aaslund L., Henriksson J., Campetella O., Frasch A.C.C., Pettersson U., Cazzulo J.J. Mol. Biochem. Parasitol. 45:345-348(1991) [PubMed: 2038364] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA] OF 295-467. Strain: Tulahuen 2.
[5]	"Self-proteolysis of the cysteine proteinase, cruzipain, from Trypanosoma cruzi gives a major fragment corresponding to its carboxy-terminal domain." Hellman U., Wernstedt C., Cazzulo J.J. Mol. Biochem. Parasitol. 44:15-21(1991) [PubMed: 2011151] [Abstract] Cited for: AUTOCATALYSIS OF C-TERMINAL. Strain: Tulahuen 2.
[6]	"Some kinetic properties of a cysteine proteinase (cruzipain) from Trypanosoma cruzi." Cazzulo J.J., Cazzulo-Franke M.C., Martinez J., Franke de Cazzulo B.M. Biochim. Biophys. Acta 1037:186-191(1990) [PubMed: 2407295] [Abstract] Cited for: SPECIFICITY. Strain: Tulahuen 2.
[7]	"Further characterization and partial amino acid sequence of a cysteine proteinase from Trypanosoma cruzi." Cazzulo J.J., Couso R., Raimondi A., Wernstedt C., Hellman U. Mol. Biochem. Parasitol. 33:33-42(1989) [PubMed: 2651912] [Abstract] Cited for: PROTEIN SEQUENCE OF 123-146 AND 304-317.
[8]	"Structural determinants of specificity in the cysteine protease cruzain." Gillmor S.A., Craik C.S., Fletterick R.J. Protein Sci. 6:1603-1611(1997) [PubMed: 9260273] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 123-337.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

M84342 Genomic DNA. Translation: AAA30181.1.
M69121 Genomic DNA. Translation: AAA30269.1.
M69121 Genomic DNA. Translation: AAA30270.1.
X54414 mRNA. Translation: CAA38278.1.
M27305 Genomic DNA. Translation: AAA30180.1.

PIR

A60667.
S16162.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1AIM	X-ray	2.00	A	123-337	[»]
1EWL	X-ray	2.00	A	123-337	[»]
1EWM	X-ray	2.00	A	123-337	[»]
1EWO	X-ray	2.10	A	123-337	[»]
1EWP	X-ray	1.75	A	123-337	[»]
1F29	X-ray	2.15	A/B/C	123-337	[»]
1F2A	X-ray	1.60	A	123-337	[»]
1F2B	X-ray	1.80	A	123-337	[»]
1F2C	X-ray	2.00	A	123-337	[»]
1ME3	X-ray	1.20	A	123-337	[»]
1ME4	X-ray	1.20	A	123-337	[»]
1U9Q	X-ray	2.30	X	123-337	[»]
2AIM	X-ray	2.20	A	123-337	[»]
2EFM	X-ray	1.50	A/C	123-337	[»]
2OZ2	X-ray	1.95	A/C	123-337	[»]

ModBase

Search...

Protein family/group databases

MEROPS

C01.075.

Enzyme and pathway databases

BRENDA

3.4.22.51. 884.

Family and domain databases

InterPro

IPR000169. Pept_cys_AS.
IPR013128. Peptidase_C1A.
IPR000668. Peptidase_C1A_C.
IPR013201. Prot_inhib_I29.
[Graphical view]

PANTHER

PTHR12411. Peptidase_C1A. 1 hit.

Pfam

PF08246. Inhibitor_I29. 1 hit.
PF00112. Peptidase_C1. 1 hit.
[Graphical view]

PRINTS

PR00705. PAPAIN.

ProDom

PD000158. Peptidase_C1. 1 hit.
[Graphical view] [Entries sharing at least one domain]

SMART

SM00645. Pept_C1. 1 hit.
[Graphical view]

PROSITE

PS00640. THIOL_PROTEASE_ASN. 1 hit.
PS00139. THIOL_PROTEASE_CYS. 1 hit.
PS00639. THIOL_PROTEASE_HIS. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

CYSP_TRYCR

Accession

Primary (citable) accession number: P25779

Entry history

Integrated into UniProtKB/Swiss-Prot:	May 1, 1992
Last sequence update:	May 1, 1992
Last modified:	June 16, 2009
This is version 82 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)