P00782 (SUBT_BACAM) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Subtilisin BPN'
EC=3.4.21.62

Alternative name(s):
Alkaline protease
Subtilisin DFE
Subtilisin Novo

Gene names

Name:

apr

Organism

Bacillus amyloliquefaciens (Bacillus velezensis)

Taxonomic identifier

1390 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Firmicutes › Bacilli › Bacillales › Bacillaceae › Bacillus › Bacillus subtilis group

Protein attributes

Sequence length	382 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	Subtilisin is an extracellular alkaline serine protease, it catalyzes the hydrolysis of proteins and peptide amides. Has a high substrate specificity to fibrin. Ref.4
Catalytic activity	Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides.
Cofactor	Binds 2 calcium ions per subunit.
Enzyme regulation	Completely inhibited by phenylmethylsulfony fluoride (PMSF) and partially inhibited by benzamidine hydrochloride, leupeptin, and pepstatin A. Ref.4
Subunit structure	Monomer. Ref.4
Subcellular location	Secreted.
Biotechnological use	Used as a detergent protease. Sold under the name Alcalase by Novozymes.
Miscellaneous	Secretion of subtilisin is associated with onset of sporulation, and many mutations which block sporulation at early stages affect expression levels of subtilisin. However, subtilisin is not necessary for normal sporulation.
Sequence similarities	Belongs to the peptidase S8 family.
Biophysicochemical properties	pH dependence: Optimum pH is 9.0. Ref.4 Temperature dependence: Optimum temperature is 48 degrees Celsius.
Sequence caution	The sequence CAA24990.1 differs from that shown. Reason: Erroneous initiation.

Ontologies

Keywords
Biological process	Sporulation
Cellular component	Secreted
Domain	Signal
Ligand	Calcium Metal-binding
Molecular function	Hydrolase Protease Serine protease
PTM	Zymogen
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological_process	fibrinolysis Inferred from direct assay Ref.4. Source: UniProtKB negative regulation of catalytic activity Inferred from electronic annotation. Source: InterPro proteolysis Inferred from direct assay Ref.4. Source: UniProtKB sporulation resulting in formation of a cellular spore Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW serine-type endopeptidase activity Inferred from direct assay Ref.4. Source: UniProtKB
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
Ica-2	Q40059	8	EBI-1033955,EBI-1040468	From a different organism.

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 30

30

By similarity

Propeptide

31 – 107

77

PRO_0000027177

Chain

108 – 382

275

Subtilisin BPN'

PRO_0000027178

Sites

Active site

139

1

Charge relay system Ref.9

Active site

171

1

Charge relay system Ref.9

Active site

328

1

Charge relay system Ref.9

Metal binding

109

1

Calcium 1

Metal binding

148

1

Calcium 1

Metal binding

182

1

Calcium 1; via carbonyl oxygen

Metal binding

184

1

Calcium 1

Metal binding

186

1

Calcium 1; via carbonyl oxygen

Metal binding

188

1

Calcium 1; via carbonyl oxygen

Metal binding

276

1

Calcium 2; via carbonyl oxygen

Metal binding

278

1

Calcium 2; via carbonyl oxygen

Metal binding

281

1

Calcium 2; via carbonyl oxygen

Natural variations

Natural variant

128

1

Y → F in strain: DC-4. Ref.4

Secondary structure

1

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382

Helix Strand Turn

Details...

Helix

113 – 117

5

Helix

120 – 126

7

Beta strand

134 – 140

7

Beta strand

146 – 148

3

Beta strand

151 – 156

6

Beta strand

158 – 160

3

Beta strand

168 – 170

3

Helix

171 – 180

10

Beta strand

183 – 188

6

Beta strand

195 – 201

7

Beta strand

207 – 209

3

Helix

211 – 223

13

Beta strand

227 – 231

5

Beta strand

233 – 236

4

Helix

240 – 251

12

Beta strand

255 – 259

5

Turn

275 – 277

3

Beta strand

281 – 287

7

Beta strand

289 – 291

3

Beta strand

305 – 308

4

Beta strand

310 – 316

7

Turn

317 – 319

3

Beta strand

320 – 324

5

Helix

327 – 344

18

Helix

350 – 358

9

Helix

367 – 370

4

Helix

377 – 380

4

Sequences

	Sequence		Length	Mass (Da)
	P00782 [UniParc]. Last modified July 21, 1986. Version 1. Checksum: ED987DAFA37B8335	FASTA	382	39,181
	10 20 30 40 50 60 MRGKKVWISL LFALALIFTM AFGSTSSAQA AGKSNGEKKY IVGFKQTMST MSAAKKKDVI 70 80 90 100 110 120 SEKGGKVQKQ FKYVDAASAT LNEKAVKELK KDPSVAYVEE DHVAHAYAQS VPYGVSQIKA 130 140 150 160 170 180 PALHSQGYTG SNVKVAVIDS GIDSSHPDLK VAGGASMVPS ETNPFQDNNS HGTHVAGTVA 190 200 210 220 230 240 ALNNSIGVLG VAPSASLYAV KVLGADGSGQ YSWIINGIEW AIANNMDVIN MSLGGPSGSA 250 260 270 280 290 300 ALKAAVDKAV ASGVVVVAAA GNEGTSGSSS TVGYPGKYPS VIAVGAVDSS NQRASFSSVG 310 320 330 340 350 360 PELDVMAPGV SIQSTLPGNK YGAYNGTSMA SPHVAGAAAL ILSKHPNWTN TQVRSSLENT 370 380 TTKLGDSFYY GKGLINVQAA AQ

References

[1]	"Genes for alkaline protease and neutral protease from Bacillus amyloliquefaciens contain a large open reading frame between the regions coding for signal sequence and mature protein." Vasantha N., Thompson L.D., Rhodes C., Banner C., Nagle J., Filpula D. J. Bacteriol. 159:811-819(1984) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: ATCC 23844 / P.
[2]	"Cloning, sequencing, and secretion of Bacillus amyloliquefaciens subtilisin in Bacillus subtilis." Wells J.A., Ferrari E., Henner D.J., Estell D.A., Chen E.Y. Nucleic Acids Res. 11:7911-7925(1983) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 2-382.
[3]	"Subtilisin BPN. VII. Isolation of cyanogen bromide peptides and the complete amino acid sequence." Markland F.S., Smith E.L. J. Biol. Chem. 242:5198-5211(1967) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 108-382.
[4]	"Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional Chinese soybean food." Peng Y., Huang Q., Zhang R.-H., Zhang Y.-Z. Comp. Biochem. Physiol. 134B:45-52(2003) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 108-131, FUNCTION, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION, VARIANT PHE-128. Strain: DC-4.
[5]	"A hydrogen-bond network at the active site of subtilisin BPN'." Alden R.A., Wright C.S., Kraut J. Philos. Trans. R. Soc. Lond., B, Biol. Sci. 257:119-124(1970) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS).
[6]	"Crystal structure at 2.6-A resolution of the complex of subtilisin BPN' with streptomyces subtilisin inhibitor." Hirono S., Akagawa H., Mitsui Y., Iitaka Y. J. Mol. Biol. 178:389-413(1984) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) OF COMPLEX WITH INHIBITOR.
[7]	"Large increases in general stability for subtilisin BPN' through incremental changes in the free energy of unfolding." Pantoliano M.W., Whitlow M., Wood J.F., Dodd S.W., Hardman K.D., Rollence M.L., Bryan P.N. Biochemistry 28:7205-7213(1989) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) OF MUTANT.
[8]	"Subtilisin BPN' at 1.6-A resolution: analysis for discrete disorder and comparison of crystal forms." Gallagher T., Oliver J., Bott R., Betzel C., Gilliland G.L. Acta Crystallogr. D 52:1125-1135(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS).
[9]	"Identification of histidine 64 in the active site of subtilisin." Markland F.S., Shaw E., Smith E.L. Proc. Natl. Acad. Sci. U.S.A. 61:1440-1447(1968) [PubMed] [Europe PMC] [Abstract] Cited for: ACTIVE SITE.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

K02496 Genomic DNA. Translation: AAB05345.1.
X00165 Genomic DNA. Translation: CAA24990.1. Different initiation.

PIR

SUBSN. B25415.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1A2Q	X-ray	1.80	A	108-382	[»]
1AK9	X-ray	1.80	A	108-382	[»]
1AQN	X-ray	1.80	A	108-382	[»]
1AU9	X-ray	1.80	A	108-382	[»]
1DUI	X-ray	2.00	A	113-382	[»]
1GNS	X-ray	1.80	A	112-382	[»]
1GNV	X-ray	1.90	A	113-382	[»]
1LW6	X-ray	1.50	E	108-382	[»]
1S01	X-ray	1.70	A	108-382	[»]
1S02	X-ray	1.90	A	108-382	[»]
1SBH	X-ray	1.80	A	108-382	[»]
1SBI	X-ray	2.20	A	108-382	[»]
1SBN	X-ray	2.10	E	108-382	[»]
1SBT	X-ray	2.50	A	108-382	[»]
1SIB	X-ray	2.40	E	108-382	[»]
1SPB	X-ray	2.00	P	37-107	[»]
			S	108-382	[»]
1ST2	X-ray	2.00	A	108-382	[»]
1SUA	X-ray	2.10	A	108-382	[»]
1SUB	X-ray	1.75	A	108-382	[»]
1SUC	X-ray	1.80	A	108-382	[»]
1SUD	X-ray	1.90	A	108-382	[»]
1SUE	X-ray	1.80	A	113-382	[»]
1SUP	X-ray	1.60	A	108-382	[»]
1TM1	X-ray	1.70	E	108-382	[»]
1TM3	X-ray	1.57	E	108-382	[»]
1TM4	X-ray	1.70	E	108-382	[»]
1TM5	X-ray	1.45	E	108-382	[»]
1TM7	X-ray	1.59	E	108-382	[»]
1TMG	X-ray	1.67	E	108-382	[»]
1TO1	X-ray	1.68	E	108-382	[»]
1TO2	X-ray	1.30	E	108-382	[»]
1UBN	X-ray	2.40	A	108-382	[»]
1V5I	X-ray	1.50	A	108-382	[»]
1Y1K	X-ray	1.56	E	108-382	[»]
1Y33	X-ray	1.80	E	108-382	[»]
1Y34	X-ray	1.55	E	108-382	[»]
1Y3B	X-ray	1.80	E	108-382	[»]
1Y3C	X-ray	1.69	E	108-382	[»]
1Y3D	X-ray	1.80	E	108-382	[»]
1Y3F	X-ray	1.72	E	108-382	[»]
1Y48	X-ray	1.84	E	108-382	[»]
1Y4A	X-ray	1.60	E	108-382	[»]
1Y4D	X-ray	2.00	E	108-382	[»]
1YJA	X-ray	1.80	A	108-382	[»]
1YJB	X-ray	1.80	A	108-382	[»]
1YJC	X-ray	1.80	A	108-382	[»]
2SBT	X-ray	2.80	A	108-382	[»]
2SIC	X-ray	1.80	E	108-382	[»]
2SNI	X-ray	2.10	E	108-382	[»]
2ST1	X-ray	1.80	A	108-382	[»]
3BGO	X-ray	1.80	S	113-382	[»]
3CNQ	X-ray	1.71	P	32-101	[»]
			S	113-382	[»]
3CO0	X-ray	1.93	P	32-101	[»]
			S	113-382	[»]
3F49	X-ray	1.70	S	113-382	[»]
3SIC	X-ray	1.80	E	108-382	[»]
5SIC	X-ray	2.20	E	108-382	[»]

ProteinModelPortal

P00782.

SMR

P00782. Positions 37-382.

ModBase

Search...

Protein-protein interaction databases

IntAct

P00782. 2 interactions.

MINT

MINT-242921.

Protein family/group databases

MEROPS

I09.001.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Family and domain databases

Gene3D

3.40.50.200. 1 hit.

InterPro

IPR010259. Inhibitor_I9.
IPR000209. Peptidase_S8/S53_dom.
IPR023827. Peptidase_S8_Asp-AS.
IPR022398. Peptidase_S8_His-AS.
IPR023828. Peptidase_S8_Ser-AS.
IPR015500. Peptidase_S8_subtilisin-rel.
IPR009020. Prot_inh_propept.
[Graphical view]

PANTHER

PTHR10795. PTHR10795. 1 hit.

Pfam

PF05922. Inhibitor_I9. 1 hit.
PF00082. Peptidase_S8. 1 hit.
[Graphical view]

PRINTS

PR00723. SUBTILISIN.

SUPFAM

SSF52743. Pept_S8_S53. 1 hit.
SSF54897. Prot_inh_propept. 1 hit.

PROSITE

PS00136. SUBTILASE_ASP. 1 hit.
PS00137. SUBTILASE_HIS. 1 hit.
PS00138. SUBTILASE_SER. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

P00782.

PMAP-CutDB

P00782.

Entry information

Entry name

SUBT_BACAM

Accession

Primary (citable) accession number: P00782
Secondary accession number(s): P83945, Q44684

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 21, 1986
Last sequence update:	July 21, 1986
Last modified:	April 3, 2013
This is version 120 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program