P65728 (PKNG_MYCTU) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Serine/threonine-protein kinase PknG
EC=2.7.11.1

Gene names

Name:	pknG
Ordered Locus Names:	Rv0410c, MT0423
ORF Names:	MTCY22G10.06c

Organism

Mycobacterium tuberculosis [Reference proteome] [HAMAP]

Taxonomic identifier

1773 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Actinobacteria › Actinobacteria › Actinobacteridae › Actinomycetales › Corynebacterineae › Mycobacteriaceae › Mycobacterium › Mycobacterium tuberculosis complex

Protein attributes

Sequence length	750 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Phosphorylates GarA. May play a role in metabolic regulation via control of the phosphorylation status of GarA. Plays a crucial role in the survival of mycobacteria within host macrophages, by blocking the intracellular degradation of mycobacteria in lysosomes. Required for intrinsic antibiotic resistance. Ref.5 Ref.6 Ref.7 Ref.8
Catalytic activity	ATP + a protein = ADP + a phosphoprotein. Ref.3 Ref.4 Ref.5 Ref.7
Enzyme regulation	Kinase activity is regulated by the redox status of the environment via the rubredoxin domain. Autophosphorylation is not essential for kinase activity, but it promotes binding to GarA. The C-terminal domain also contributes to the regulation of activity. Inhibited by a specific small molecular-weight inhibitor, the tetrahydrobenzothiophene AX20017. Ref.5 Ref.7 Ref.8
Subunit structure	Homodimer. Interacts with the FHA domain of GarA. Ref.5 Ref.8
Subcellular location	Cytoplasm. Cell membrane. Note: Also detected in growth media, suggesting that it can be translocated into host macrophages under specific conditions. Ref.3 Ref.4
Domain	Contains an N-terminal rubredoxin domain, a central kinase domain and a C-terminal TPR domain. Ref.7 Ref.8
Post-translational modification	Autophosphorylated. In vitro, incorporates up to four phosphate groups on Thr-23, Thr-32 and Thr-63 and/or Thr-64 and/or Ser-65. In vivo, is probably phosphorylated only on Thr-63. Ref.3 Ref.4 Ref.5 Ref.7
Disruption phenotype	Disruption causes delayed mortality in mice. Mutant accumulates glutamate and glutamine. Ref.3
Sequence similarities	Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Contains 1 protein kinase domain. Contains 1 TPR repeat.
Biophysicochemical properties	Kinetic parameters: KM=2122 nM for GarA Ref.7
Sequence caution	The sequence AAK44647.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened.

Ontologies

Keywords
Biological process	Antibiotic resistance Virulence
Cellular component	Cell membrane Cytoplasm Membrane
Domain	TPR repeat
Ligand	ATP-binding Nucleotide-binding
Molecular function	Kinase Serine/threonine-protein kinase Transferase
PTM	Phosphoprotein
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	avoidance of host defenses Inferred from mutant phenotype PubMed 15155913. Source: MTBBASE glutamate catabolic process Inferred from mutant phenotype Ref.3. Source: MTBBASE glutamine catabolic process Inferred from mutant phenotype Ref.3. Source: MTBBASE growth Inferred from mutant phenotype PubMed 12657046. Source: MTBBASE modulation by symbiont of host phagocytosis Inferred from mutant phenotype PubMed 19447903. Source: MTBBASE modulation by symbiont of host signal transduction pathway Inferred from mutant phenotype PubMed 15155913. Source: MTBBASE negative regulation of cytolysis by symbiont of host cells Inferred from mutant phenotype PubMed 20030858. Source: MTBBASE pathogenesis Inferred from mutant phenotype PubMed 15155913Ref.3. Source: MTBBASE positive regulation by symbiont of defense-related host reactive oxygen species production Inferred from mutant phenotype PubMed 19447903. Source: MTBBASE protein autophosphorylation Inferred from direct assay Ref.3Ref.5PubMed 19447903. Source: MTBBASE response to antibiotic Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	cytosol Inferred from direct assay Ref.3. Source: MTBBASE plasma membrane Inferred from direct assay Ref.3PubMed 15525680. Source: MTBBASE
Molecular_function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW protein serine/threonine kinase activity Inferred from direct assay Ref.4Ref.5. Source: MTBBASE
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
garA	P64897	4	EBI-6405537,EBI-6405522

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 750

750

Serine/threonine-protein kinase PknG

PRO_0000171216

Regions

Domain

151 – 396

246

Protein kinase

Repeat

536 – 569

34

TPR

Nucleotide binding

157 – 165

9

ATP By similarity

Sites

Active site

276

1

Proton acceptor By similarity

Binding site

181

1

ATP By similarity

Amino acid modifications

Modified residue

63

1

Phosphothreonine Ref.7

Experimental info

Mutagenesis

63

1

T → A: Lack of phosphorylation. Does not affect kinase activity. Ref.7

Mutagenesis

87

1

I → S: Decreases inhibition by AX20017; when associated with S-92. Ref.8

Mutagenesis

92

1

A → S: Decreases inhibition by AX20017; when associated with S-87. Ref.8

Mutagenesis

106

1

C → A: Decrease in activity; when associated with A-109; A-128 and A-131. Ref.7 Ref.8

Mutagenesis

106

1

C → S: Lack of activity; when associated with S-109; S-128 and S-131. Ref.7 Ref.8

Mutagenesis

109

1

C → A: Decrease in activity; when associated with A-106; A-128 and A-131. Ref.7 Ref.8

Mutagenesis

109

1

C → S: Lack of activity; when associated with S-106; S-128 and S-131. Ref.7 Ref.8

Mutagenesis

128

1

C → A: Decrease in activity; when associated with A-106; A-109 and A-131. Ref.7 Ref.8

Mutagenesis

128

1

C → S: Lack of activity; when associated with S-106; S-109 and S-131. Ref.7 Ref.8

Mutagenesis

131

1

C → A: Decrease in activity; when associated with A-106; A-109 and A-128. Ref.7 Ref.8

Mutagenesis

131

1

C → S: Lack of activity; when associated with S-106; S-109 and S-128. Ref.7 Ref.8

Mutagenesis

181

1

K → M: Lack of activity. Ref.4 Ref.7

Mutagenesis

309

1

T → A, D or E: Lack of activity. Ref.7

Mutagenesis

309

1

T → S: Decrease in activity. Ref.7

Secondary structure

1

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750

Helix Strand Turn

Details...

Turn

75 – 78

4

Beta strand

79 – 82

4

Helix

89 – 92

4

Helix

101 – 103

3

Turn

107 – 109

3

Beta strand

124 – 127

4

Turn

129 – 131

3

Beta strand

134 – 136

3

Turn

148 – 150

3

Beta strand

151 – 160

10

Beta strand

163 – 170

8

Helix

171 – 173

3

Beta strand

177 – 183

7

Helix

189 – 199

11

Helix

200 – 204

5

Beta strand

213 – 220

8

Beta strand

226 – 233

8

Helix

250 – 269

20

Helix

279 – 281

3

Beta strand

282 – 284

3

Beta strand

289 – 291

3

Turn

310 – 312

3

Helix

317 – 320

4

Helix

324 – 339

16

Helix

358 – 362

5

Helix

364 – 373

10

Helix

378 – 380

3

Helix

385 – 404

20

Beta strand

413 – 415

3

Beta strand

423 – 425

3

Helix

426 – 429

4

Helix

430 – 432

3

Helix

433 – 436

4

Helix

446 – 452

7

Helix

465 – 470

6

Turn

471 – 473

3

Helix

476 – 487

12

Helix

503 – 515

13

Helix

518 – 532

15

Helix

536 – 549

14

Helix

552 – 565

14

Helix

571 – 583

13

Helix

591 – 598

8

Helix

603 – 615

13

Helix

619 – 627

9

Helix

636 – 646

11

Helix

658 – 669

12

Helix

678 – 693

16

Beta strand

699 – 703

5

Beta strand

706 – 709

4

Helix

710 – 727

18

Helix

731 – 744

14

Sequences

	Sequence		Length	Mass (Da)
	P65728 [UniParc]. Last modified October 11, 2004. Version 1. Checksum: CFC6E54DBE19569F	FASTA	750	81,577
	10 20 30 40 50 60 MAKASETERS GPGTQPADAQ TATSATVRPL STQAVFRPDF GDEDNFPHPT LGPDTEPQDR 70 80 90 100 110 120 MATTSRVRPP VRRLGGGLVE IPRAPDIDPL EALMTNPVVP ESKRFCWNCG RPVGRSDSET 130 140 150 160 170 180 KGASEGWCPY CGSPYSFLPQ LNPGDIVAGQ YEVKGCIAHG GLGWIYLALD RNVNGRPVVL 190 200 210 220 230 240 KGLVHSGDAE AQAMAMAERQ FLAEVVHPSI VQIFNFVEHT DRHGDPVGYI VMEYVGGQSL 250 260 270 280 290 300 KRSKGQKLPV AEAIAYLLEI LPALSYLHSI GLVYNDLKPE NIMLTEEQLK LIDLGAVSRI 310 320 330 340 350 360 NSFGYLYGTP GFQAPEIVRT GPTVATDIYT VGRTLAALTL DLPTRNGRYV DGLPEDDPVL 370 380 390 400 410 420 KTYDSYGRLL RRAIDPDPRQ RFTTAEEMSA QLTGVLREVV AQDTGVPRPG LSTIFSPSRS 430 440 450 460 470 480 TFGVDLLVAH TDVYLDGQVH AEKLTANEIV TALSVPLVDP TDVAASVLQA TVLSQPVQTL 490 500 510 520 530 540 DSLRAARHGA LDADGVDFSE SVELPLMEVR ALLDLGDVAK ATRKLDDLAE RVGWRWRLVW 550 560 570 580 590 600 YRAVAELLTG DYDSATKHFT EVLDTFPGEL APKLALAATA ELAGNTDEHK FYQTVWSTND 610 620 630 640 650 660 GVISAAFGLA RARSAEGDRV GAVRTLDEVP PTSRHFTTAR LTSAVTLLSG RSTSEVTEEQ 670 680 690 700 710 720 IRDAARRVEA LPPTEPRVLQ IRALVLGGAL DWLKDNKAST NHILGFPFTS HGLRLGVEAS 730 740 750 LRSLARVAPT QRHRYTLVDM ANKVRPTSTF

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence." Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E., Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K., Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K. , Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K., Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J., Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S., Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S., Barrell B.G. Nature 393:537-544(1998) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: ATCC 25618 / H37Rv.
[2]	"Whole-genome comparison of Mycobacterium tuberculosis clinical and laboratory strains." Fleischmann R.D., Alland D., Eisen J.A., Carpenter L., White O., Peterson J.D., DeBoy R.T., Dodson R.J., Gwinn M.L., Haft D.H., Hickey E.K., Kolonay J.F., Nelson W.C., Umayam L.A., Ermolaeva M.D., Salzberg S.L., Delcher A., Utterback T.R. , Weidman J.F., Khouri H.M., Gill J., Mikula A., Bishai W., Jacobs W.R. Jr., Venter J.C., Fraser C.M. J. Bacteriol. 184:5479-5490(2002) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: CDC 1551 / Oshkosh.
[3]	"The Mycobacterium tuberculosis protein serine/threonine kinase PknG is linked to cellular glutamate/glutamine levels and is important for growth in vivo." Cowley S., Ko M., Pick N., Chow R., Downing K.J., Gordhan B.G., Betts J.C., Mizrahi V., Smith D.A., Stokes R.W., Av-Gay Y. Mol. Microbiol. 52:1691-1702(2004) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-10, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, AUTOPHOSPHORYLATION, DISRUPTION PHENOTYPE. Strain: ATCC 25618 / H37Rv.
[4]	"Serine/threonine protein kinases PknF and PknG of Mycobacterium tuberculosis: characterization and localization." Koul A., Choidas A., Tyagi A.K., Drlica K., Singh Y., Ullrich A. Microbiology 147:2307-2314(2001) [PubMed] [Europe PMC] [Abstract] Cited for: CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, AUTOPHOSPHORYLATION, MUTAGENESIS OF LYS-181. Strain: ATCC 25618 / H37Rv.
[5]	"Regulation of glutamate metabolism by protein kinases in mycobacteria." O'Hare H.M., Duran R., Cervenansky C., Bellinzoni M., Wehenkel A.M., Pritsch O., Obal G., Baumgartner J., Vialaret J., Johnsson K., Alzari P.M. Mol. Microbiol. 70:1408-1423(2008) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, INTERACTION WITH GARA, AUTOPHOSPHORYLATION. Strain: ATCC 25618 / H37Rv.
[6]	"Protein kinase G is required for intrinsic antibiotic resistance in mycobacteria." Wolff K.A., Nguyen H.T., Cartabuke R.H., Singh A., Ogwang S., Nguyen L. Antimicrob. Agents Chemother. 53:3515-3519(2009) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN ANTIBIOTIC RESISTANCE. Strain: ATCC 25618 / H37Rv.
[7]	"Key residues in Mycobacterium tuberculosis protein kinase G play a role in regulating kinase activity and survival in the host." Tiwari D., Singh R.K., Goswami K., Verma S.K., Prakash B., Nandicoori V.K. J. Biol. Chem. 284:27467-27479(2009) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION, DOMAIN, AUTOPHOSPHORYLATION AT THR-63, MUTAGENESIS OF THR-63; CYS-106; CYS-109; CYS-128; CYS-131; LYS-181 AND THR-309. Strain: ATCC 25618 / H37Rv.
[8]	"Structural basis for the specific inhibition of protein kinase G, a virulence factor of Mycobacterium tuberculosis." Scherr N., Honnappa S., Kunz G., Mueller P., Jayachandran R., Winkler F., Pieters J., Steinmetz M.O. Proc. Natl. Acad. Sci. U.S.A. 104:12151-12156(2007) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 74-750 IN COMPLEX WITH INHIBITOR AX20017, FUNCTION, ENZYME REGULATION, SUBUNIT, DOMAIN, MUTAGENESIS OF ILE-87; ALA-92; CYS-106; CYS-109; CYS-128 AND CYS-131.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

BX842573 Genomic DNA. Translation: CAB06580.1.
AE000516 Genomic DNA. Translation: AAK44647.1. Different initiation.
AL123456 Genomic DNA. Translation: CCP43141.1.

PIR

H70628.

RefSeq

NP_214924.1. NC_000962.3.
NP_334833.1. NC_002755.2.
YP_006513736.1. NC_018143.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2PZI	X-ray	2.40	A/B	74-750	[»]

ProteinModelPortal

P65728.

SMR

P65728. Positions 74-746.

ModBase

Search...

Protein-protein interaction databases

IntAct

P65728. 1 interaction.

STRING

83332.Rv0410c.

PTM databases

PhosSite

P0904617.

Proteomic databases

PRIDE

P65728.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAK44647; AAK44647; MT0423.

GeneID

13318277.
886397.
923697.

KEGG

mtc:MT0423.
mtu:Rv0410c.
mtv:RVBD_0410c.

PATRIC

18122648. VBIMycTub22151_0458.

Organism-specific databases

TubercuList

Rv0410c.

Phylogenomic databases

eggNOG

COG0515.

HOGENOM

HOG000049016.

KO

K14949.

OMA

HRYALVD.

ProtClustDB

CLSK790502.

Family and domain databases

InterPro

IPR011009. Kinase-like_dom.
IPR000719. Prot_kinase_cat_dom.
IPR008271. Ser/Thr_kinase_AS.
[Graphical view]

Pfam

PF00069. Pkinase. 1 hit.
[Graphical view]

SUPFAM

SSF56112. Kinase_like. 1 hit.

PROSITE

PS00107. PROTEIN_KINASE_ATP. False negative.
PS50011. PROTEIN_KINASE_DOM. 1 hit.
PS00108. PROTEIN_KINASE_ST. 1 hit.
PS50005. TPR. False negative.
PS50293. TPR_REGION. False negative.
[Graphical view]

ProtoNet

Search...

Other

ChEMBL

CHEMBL6070.

EvolutionaryTrace

P65728.

Entry information

Entry name

PKNG_MYCTU

Accession

Primary (citable) accession number: P65728
Secondary accession number(s): L0T3M0, P96256

Entry history

Integrated into UniProtKB/Swiss-Prot:	October 11, 2004
Last sequence update:	October 11, 2004
Last modified:	May 1, 2013
This is version 65 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program