Reviewed, UniProtKB/Swiss-Prot Q06320 (CWLC_BACSU)

Names and origin

Protein names

Recommended name:
    Sporulation-specific N-acetylmuramoyl-L-alanine amidase
    EC=3.5.1.28
Alternative name(s):
    Cell wall hydrolase
    Autolysin

Gene names

Name:	cwlC
Ordered Locus Names:	BSU17410

Organism

Bacillus subtilis [Complete proteome] [HAMAP]

Taxonomic identifier

1423 [NCBI]

Taxonomic lineage

Bacteria › Firmicutes › Bacillales › Bacillaceae › Bacillus

Protein attributes

Sequence length	255 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	Autolysins are involved in some important biological processes such as cell separation, cell-wall turnover, competence for genetic transformation, formation of the flagella - in particular of its basal body - and sporulation. CwlC is able to hydrolyze type A cell walls such as B.subtilis. Its main function is to lyze the mother cell wall peptidoglycan, playing a role during sporulation. Ref.4 Ref.6
Catalytic activity	Hydrolyzes the link between N-acetylmuramoyl residues and L-amino acid residues in certain cell-wall glycopeptides.
Cofactor	Zinc Probable. Ref.7
Enzyme regulation	Inhibited by EDTA. Ref.6
Subcellular location	Secreted › cell wall. Note: Is associated with the mother cell wall of sporulating cells. Ref.4
Induction	Induced at 6 to 7 hours after sporulation. Ref.6 Ref.1
Domain	Contains an N-terminal catalytic domain and two C-terminal tandem repeat sequences that play an important role in peptidoglycan binding. Ref.8
Sequence similarities	Belongs to the N-acetylmuramoyl-L-alanine amidase 3 family.
Biophysicochemical properties	pH dependence: Optimum pH is 8.5-9.5. Temperature dependence: Optimum temperature is 60 degrees Celsius.

Ontologies

Keywords
Biological process	Cell wall biogenesis/degradation Competence Sporulation
Cellular component	Cell wall Secreted
Domain	Repeat
Ligand	Metal-binding Zinc
Molecular function	Hydrolase
Technical term	3D-structure Complete proteome Direct protein sequencing
Gene Ontology (GO)
Biological process	cellular cell wall organization Inferred from electronic annotation. Source: UniProtKB-KW establishment of competence for transformation Inferred from electronic annotation. Source: UniProtKB-KW peptidoglycan catabolic process Inferred from electronic annotation. Source: InterPro sporulation resulting in formation of a cellular spore Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	cell wall Inferred from electronic annotation. Source: UniProtKB-SubCell extracellular region Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	N-acetylmuramoyl-L-alanine amidase activity Inferred from electronic annotation. Source: EC 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

Initiator methionine

1

1

Removed Ref.4

Chain

2 – 255

254

Sporulation-specific N-acetylmuramoyl-L-alanine amidase

PRO_0000164418

Regions

Repeat

184 – 219

36

1

Repeat

220 – 255

36

2

Region

184 – 255

72

2 X 35 AA approximate tandem repeats

Sites

Active site

141

1

Potential

Metal binding

10

1

Zinc Potential

Metal binding

24

1

Zinc Potential

Metal binding

79

1

Zinc Potential

Experimental info

Mutagenesis

10

1

H → Q: 23% of wild-type activity. Ref.7

Mutagenesis

24

1

E → A, Q or S: Loss of activity. Ref.7

Mutagenesis

55

1

D → V: Loss of activity. Ref.7

Mutagenesis

79

1

H → L: Loss of activity. Ref.7

Mutagenesis

79

1

H → Q: 15% of wild-type activity. Ref.7

Mutagenesis

81

1

N → I: Loss of activity. Ref.7

Mutagenesis

141

1

E → Q or V: Loss of activity. Ref.7

Sequence conflict

158

1

S → T in CAA92813. Ref.5

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

255

Helix Strand Turn

Details...

Beta strand

185 – 194

10

Helix

196 – 209

14

Beta strand

213 – 218

6

Beta strand

221 – 231

11

Helix

232 – 245

14

Beta strand

250 – 254

5

Sequences

	Sequence		Length	Mass (Da)
	Q06320-1 [UniParc]. Last modified February 1, 1995. Version 1. Checksum: 30C950116784FBC1	FASTA	255	27,146
	10 20 30 40 50 60 MVKIFIDPGH GGSDPGATGN GLQEKTLTLQ IALALRTILT NEYEGVSLLL SRTSDQYVSL 70 80 90 100 110 120 NDRTNAANNW GADFFLSIHV NSGGGTGFES YIYPDVGAPT TTYQSTIHSE VIQAVDFADR 130 140 150 160 170 180 GKKTANFHVL RESAMPALLT ENGFIDTVSD ANKLKTSSFI QSLARGHANG LEQAFNLKKT 190 200 210 220 230 240 SSSGLYKVQI GAFKVKANAD SLASNAEAKG FDSIVLLKDG LYKVQIGAFS SKDNADTLAA 250 RAKNAGFDAI VILES

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Molecular cloning of a sporulation-specific cell wall hydrolase gene of Bacillus subtilis." Kuroda A., Asami Y., Sekiguchi J. J. Bacteriol. 175:6260-6268(1993) [PubMed: 8407798] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], INDUCTION. Strain: 168.
[2]	"The complete genome sequence of the Gram-positive bacterium Bacillus subtilis." Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V., Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R., Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S., Bruschi C.V. , Caldwell B., Capuano V., Carter N.M., Choi S.-K., Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F., Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D., Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M., Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P., Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K., Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S., Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y., Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G., Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J., Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C., Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S., Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B., Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S., Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M., Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y., Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J., Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A., Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M., Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S., Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E., Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K., Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E., Yoshikawa H., Danchin A. Nature 390:249-256(1997) [PubMed: 9384377] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: 168.
[3]	"From a consortium sequence to a unified sequence: the Bacillus subtilis 168 reference genome a decade later." Barbe V., Cruveiller S., Kunst F., Lenoble P., Meurice G., Sekowska A., Vallenet D., Wang T., Moszer I., Medigue C., Danchin A. Microbiology 155:1758-1775(2009) [PubMed: 19383706] [Abstract] Cited for: SEQUENCE REVISION TO 158.
[4]	"Characterization of the involvement of two compensatory autolysins in mother cell lysis during sporulation of Bacillus subtilis 168." Smith T.J., Foster S.J. J. Bacteriol. 177:3855-3862(1995) [PubMed: 7601853] [Abstract] Cited for: PROTEIN SEQUENCE OF 2-11, FUNCTION, CHARACTERIZATION, SUBCELLULAR LOCATION. Strain: 168.
[5]	Scotti C., Valbuzzi A., Perego M., Galizzi A., Albertini A.M. Submitted (JAN-1996) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 115-255. Strain: 168.
[6]	"Overexpression, purification, and characterization of Bacillus subtilis N-acetylmuramoyl-L-alanine amidase CwlC." Shida T., Hattori H., Ise F., Sekiguchi J. Biosci. Biotechnol. Biochem. 64:1522-1525(2000) [PubMed: 10945275] [Abstract] Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION.
[7]	"Mutational analysis of catalytic sites of the cell wall lytic N-acetylmuramoyl-L-alanine amidases CwlC and CwlV." Shida T., Hattori H., Ise F., Sekiguchi J. J. Biol. Chem. 276:28140-28146(2001) [PubMed: 11375403] [Abstract] Cited for: COFACTOR, MUTAGENESIS OF HIS-10; GLU-24; ASP-55; HIS-79; ASN-81 AND GLU-141.
[8]	"Solution structure of the peptidoglycan binding domain of Bacillus subtilis cell wall lytic enzyme CwlC: characterization of the sporulation-related repeats by NMR." Mishima M., Shida T., Yabuki K., Kato K., Sekiguchi J., Kojima C. Biochemistry 44:10153-10163(2005) [PubMed: 16042392] [Abstract] Cited for: STRUCTURE BY NMR OF 177-255, DOMAIN.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

D14666 Genomic DNA. Translation: BAA03500.1.
AL009126 Genomic DNA. Translation: CAB13625.2.
Z68500 Genomic DNA. Translation: CAA92813.1.

PIR

A36935.

RefSeq

NP_389623.2.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1X60	NMR	-	A	177-255	[»]

SMR

Q06320. Positions 1-178.

ModBase

Search...

Genome annotation databases

GeneID

940088.

GenomeReviews

Gene locus BSU17410 in contig AL009126_GR.

KEGG

bsu:BSU17410.

Organism-specific databases

SubtiList

BG10825. cwlC. [Micado]

CMR

Search...

Phylogenomic databases

HOGENOM

HBG656726.

Enzyme and pathway databases

BRENDA

3.5.1.28. 150.

Family and domain databases

InterPro

IPR002508. CW_Hdrlase/autolysin_cat.
IPR007730. Sporulation_related_dom.
[Graphical view]

Gene3D

G3DSA:3.40.630.40. Cell_wall_OHase/autolysin_cat. 1 hit.

Pfam

PF01520. Amidase_3. 1 hit.
PF05036. SPOR. 1 hit.
[Graphical view]

SMART

SM00646. Ami_3. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

CWLC_BACSU

Accession

Primary (citable) accession number: Q06320

Entry history

Integrated into UniProtKB/Swiss-Prot:	February 1, 1995
Last sequence update:	February 1, 1995
Last modified:	January 19, 2010
This is version 67 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

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