Reviewed, UniProtKB/Swiss-Prot P00800 (THER_BACTH)

Names and origin

Protein names

Recommended name:
    Thermolysin
    EC=3.4.24.27
Alternative name(s):
    Thermostable neutral proteinase

Gene names

Name:

npr

Organism

Bacillus thermoproteolyticus

Taxonomic identifier

1427 [NCBI]

Taxonomic lineage

Bacteria › Firmicutes › Bacillales › Bacillaceae › Bacillus

Protein attributes

Sequence length	548 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	Extracellular zinc metalloprotease.
Catalytic activity	Preferential cleavage: Xaa-|-Leu > Xaa-|-Phe.
Cofactor	Binds 4 calcium ions per subunit. Binds 1 zinc ion per subunit.
Subcellular location	Secreted.
Sequence similarities	Belongs to the peptidase M4 family.
biophysicochemical properties	Temperature dependence: Thermostable.

Ontologies

Keywords
Cellular component	Secreted
Domain	Signal
Ligand	Calcium Metal-binding Zinc
Molecular function	Hydrolase Metalloprotease Protease
PTM	Zymogen
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological process	proteolysis Inferred from electronic annotation. Source: InterPro
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	calcium ion binding Inferred from electronic annotation. Source: UniProtKB-KW metalloendopeptidase activity Inferred from electronic annotation. Source: InterPro zinc ion binding Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 28

28

By similarity

Propeptide

29 – 232

204

Activation peptide Ref.2 Ref.3

PRO_0000028592

Chain

233 – 548

316

Thermolysin

PRO_0000028593

Sites

Active site

375

1

Ref.4

Active site

463

1

Proton donor Ref.4

Metal binding

289

1

Calcium 1

Metal binding

291

1

Calcium 1

Metal binding

293

1

Calcium 1; via carbonyl oxygen

Metal binding

370

1

Calcium 2

Metal binding

374

1

Zinc; catalytic

Metal binding

378

1

Zinc; catalytic

Metal binding

398

1

Zinc; catalytic

Metal binding

409

1

Calcium 2

Metal binding

409

1

Calcium 3

Metal binding

415

1

Calcium 3; via carbonyl oxygen

Metal binding

417

1

Calcium 2

Metal binding

417

1

Calcium 3

Metal binding

419

1

Calcium 2; via carbonyl oxygen

Metal binding

422

1

Calcium 2

Metal binding

422

1

Calcium 3

Metal binding

425

1

Calcium 4; via carbonyl oxygen

Metal binding

426

1

Calcium 4

Metal binding

429

1

Calcium 4; via carbonyl oxygen

Metal binding

432

1

Calcium 4

Experimental info

Sequence conflict

269

1

N → D AA sequence Ref.2

Sequence conflict

351

1

Q → E AA sequence Ref.2

Sequence conflict

400

1

I → M in CAA54291. Ref.1

Sequence conflict

409

1

E → K in CAA54291. Ref.1

Secondary structure

1

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548

Helix Strand Turn

Details...

Beta strand

236 – 243

8

Beta strand

249 – 264

16

Beta strand

271 – 275

5

Beta strand

280 – 283

4

Beta strand

288 – 294

7

Helix

297 – 299

3

Helix

300 – 320

21

Beta strand

332 – 337

6

Beta strand

339 – 341

3

Beta strand

345 – 347

3

Beta strand

349 – 354

6

Beta strand

359 – 363

5

Helix

365 – 367

3

Helix

369 – 383

15

Helix

391 – 411

21

Beta strand

418 – 421

4

Turn

422 – 424

3

Beta strand

434 – 438

5

Helix

440 – 443

4

Helix

449 – 451

3

Helix

457 – 461

5

Turn

462 – 465

4

Helix

466 – 478

13

Beta strand

480 – 482

3

Beta strand

485 – 487

3

Helix

492 – 505

14

Helix

513 – 528

16

Helix

533 – 545

13

Sequences

	Sequence		Length	Mass (Da)
	P00800-1 [UniParc]. Last modified April 18, 2006. Version 3. Checksum: 1D6ED3A545F045C8	FASTA	548	60,104
	10 20 30 40 50 60 MKMKMKLASF GLAAGLAAQV FLPYNALAST EHVTWNQQFQ TPQFISGDLL KVNGTSPEEL 70 80 90 100 110 120 VYQYVEKNEN KFKFHENAKD TLQLKEKKND NLGFTFMRFQ QTYKGIPVFG AVVTSHVKDG 130 140 150 160 170 180 TLTALSGTLI PNLDTKGSLK SGKKLSEKQA RDIAEKDLVA NVTKEVPEYE QGKDTEFVVY 190 200 210 220 230 240 VNGDEASLAY VVNLNFLTPE PGNWLYIIDA VDGKILNKFN QLDAAKPGDV KSITGTSTVG 250 260 270 280 290 300 VGRGVLGDQK NINTTYSTYY YLQDNTRGNG IFTYDAKYRT TLPGSLWADA DNQFFASYDA 310 320 330 340 350 360 PAVDAHYYAG VTYDYYKNVH NRLSYDGNNA AIRSSVHYSQ GYNNAFWNGS QMVYGDGDGQ 370 380 390 400 410 420 TFIPLSGGID VVAHELTHAV TDYTAGLIYQ NESGAINEAI SDIFGTLVEF YANKNPDWEI 430 440 450 460 470 480 GEDVYTPGIS GDSLRSMSDP AKYGDPDHYS KRYTGTQDNG GVHINSGIIN KAAYLISQGG 490 500 510 520 530 540 THYGVSVVGI GRDKLGKIFY RALTQYLTPT SNFSQLRAAA VQSATDLYGS TSQEVASVKQ AFDAVGVK

References

[1]	"Cloning and expression in Bacillus subtilis of the npr gene from Bacillus thermoproteolyticus Rokko coding for the thermostable metalloprotease thermolysin." O'Donohue M.J., Roques B.P., Beaumont A. Biochem. J. 300:599-603(1994) [PubMed: 8002967] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: Rokko.
[2]	"Amino-acid sequence of thermolysin." Titani K., Hermodson M.A., Ericsson L.H., Walsh K.A., Neurath H. Nature New Biol. 238:35-37(1972) Cited for: PROTEIN SEQUENCE OF 233-548.
[3]	"Amino acid sequence of thermolysin. Isolation and characterization of the fragments obtained by cleavage with cyanogen bromide." Titani K., Hermodson M.A., Ericsson L.H., Walsh K.A., Neurath H. Biochemistry 11:2427-2435(1972) [PubMed: 5040648] [Abstract] Cited for: PROTEIN SEQUENCE OF 233-268; 352-400 AND 438-477.
[4]	"Evidence of an essential histidine residue in thermolysin." Burstein Y., Walsh K.A., Neurath H. Biochemistry 13:205-210(1974) [PubMed: 4808703] [Abstract] Cited for: ACTIVE SITE.
[5]	"Structure of thermolysin refined at 1.6-A resolution." Holmes M.A., Matthews B.W. J. Mol. Biol. 160:623-639(1982) [PubMed: 7175940] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS).
[6]	"The conformation of thermolysin." Matthews B.W., Weaver L.H., Kester W.R. J. Biol. Chem. 249:8030-8044(1974) [PubMed: 4214815] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS).
[7]	"NMR solution structure of the C-terminal fragment 255-316 of thermolysin: a dimer formed by subunits having the native structure." Rico M., Jimenez M.A., Gonzalez C., de Filippis V., Fontana A. Biochemistry 33:14834-14847(1994) [PubMed: 7993910] [Abstract] Cited for: STRUCTURE BY NMR OF 487-548.
[8]	"NMR solution structure of the 205-316 C-terminal fragment of thermolysin. An example of dimerization coupled to partial unfolding." Conejero-Lara F., Gonzalez C., Jimenez M.A., Padmanabhan S., Mateo P.L., Rico M. Biochemistry 36:11975-11983(1997) [PubMed: 9305992] [Abstract] Cited for: STRUCTURE BY NMR OF 437-548.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

X76986 Genomic DNA. Translation: CAA54291.1.

PIR

HYBST. I40579.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1FJ3	X-ray	2.00	A	233-548	[»]
1FJO	X-ray	2.00	A	233-548	[»]
1FJQ	X-ray	1.70	A	233-548	[»]
1FJT	X-ray	2.20	A	233-548	[»]
1FJU	X-ray	2.00	A	233-548	[»]
1FJV	X-ray	2.00	A	233-548	[»]
1FJW	X-ray	1.90	A	233-548	[»]
1GXW	X-ray	2.18	A	233-548	[»]
1HYT	X-ray	1.70	A	233-548	[»]
1KEI	X-ray	1.60	A	233-548	[»]
1KJO	X-ray	1.60	A	233-548	[»]
1KJP	X-ray	1.60	A	233-548	[»]
1KKK	X-ray	1.60	A	233-548	[»]
1KL6	X-ray	1.80	A	233-548	[»]
1KR6	X-ray	1.80	A	233-548	[»]
1KRO	X-ray	1.70	A	233-548	[»]
1KS7	X-ray	1.70	A	233-548	[»]
1KTO	X-ray	1.90	A	233-548	[»]
1L3F	X-ray	2.30	E	233-548	[»]
1LNA	X-ray	1.90	E	233-548	[»]
1LNB	X-ray	1.80	E	233-548	[»]
1LNC	X-ray	1.80	E	233-548	[»]
1LND	X-ray	1.70	E	233-548	[»]
1LNE	X-ray	1.70	E	233-548	[»]
1LNF	X-ray	1.70	E	233-548	[»]
1OS0	X-ray	2.10	A	233-548	[»]
1PE5	X-ray	1.70	A	233-548	[»]
1PE7	X-ray	1.82	A	233-548	[»]
1PE8	X-ray	1.80	A	233-548	[»]
1QF0	X-ray	2.20	A	233-548	[»]
1QF1	X-ray	2.00	A	233-548	[»]
1QF2	X-ray	2.06	A	233-548	[»]
1THL	X-ray	1.70	A	233-548	[»]
1TLI	X-ray	2.05	A	233-548	[»]
1TLP	X-ray	2.30	E	233-548	[»]
1TLX	X-ray	2.10	A	233-548	[»]
1TMN	X-ray	1.90	E	233-548	[»]
1TRL	NMR	-	A/B	487-548	[»]
1Y3G	X-ray	2.10	E	233-548	[»]
1Z9G	X-ray	1.70	E	233-548	[»]
1ZDP	X-ray	1.70	E	233-548	[»]
2A7G	X-ray	1.85	E	233-548	[»]
2G4Z	X-ray	1.98	A	233-548	[»]
2TLI	X-ray	1.95	A	233-548	[»]
2TLX	X-ray	1.65	A	233-548	[»]
2TMN	X-ray	1.60	E	233-548	[»]
3TLI	X-ray	1.95	A	233-548	[»]
3TMN	X-ray	1.70	E	233-548	[»]
4TLI	X-ray	1.95	A	233-548	[»]
4TLN	X-ray	2.30	A	233-548	[»]
4TMN	X-ray	1.70	E	233-548	[»]
5TLI	X-ray	2.10	A	233-548	[»]
5TLN	X-ray	2.30	A	233-548	[»]
5TMN	X-ray	1.60	E	233-548	[»]
6TLI	X-ray	2.10	A	233-548	[»]
6TMN	X-ray	1.60	E	233-548	[»]
7TLI	X-ray	1.95	A	233-548	[»]
7TLN	X-ray	2.30	A	233-548	[»]
8TLI	X-ray	2.20	A	233-548	[»]
8TLN	X-ray	1.60	E	233-548	[»]

ModBase

Search...

Protein family/group databases

MEROPS

M04.001.

Enzyme and pathway databases

BRENDA

3.4.24.27. 19020.

Family and domain databases

InterPro

IPR005075. Pept_M4_propep_PepSY.
IPR006025. Pept_M_Zn_BS.
IPR013856. Peptidase_M4.
IPR001570. Peptidase_M4_C.
IPR011096. Propep_M4_M36.
[Graphical view]

Gene3D

G3DSA:3.10.170.10. Peptidase_M4. 1 hit.
G3DSA:1.10.390.10. Peptidase_M4/M36. 1 hit.

Pfam

PF07504. FTP. 1 hit.
PF03413. PepSY. 1 hit.
PF01447. Peptidase_M4. 1 hit.
PF02868. Peptidase_M4_C. 1 hit.
[Graphical view]

PRINTS

PR00730. THERMOLYSIN.

PROSITE

PS00142. ZINC_PROTEASE. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

THER_BACTH

Accession

Primary (citable) accession number: P00800
Secondary accession number(s): Q45779

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 21, 1986
Last sequence update:	April 18, 2006
Last modified:	June 16, 2009
This is version 93 of the entry and version 3 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)