P80197 (AFK_PHYPO) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names	Recommended name: Actin-fragmin kinase Short name=AFK EC=2.7.11.1
Organism	Physarum polycephalum (Slime mold)
Taxonomic identifier	5791 [NCBI]
Taxonomic lineage	Eukaryota › Amoebozoa › Mycetozoa › Myxogastria › Myxogastromycetidae › Physariida › Physaraceae › Physarum

Protein attributes

Sequence length	737 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Has a role in the regulation of microfilament formation. Phosphorylates the actin-fragmin complex on threonine residues, in vitro. Ref.1 Ref.4 Ref.5
Catalytic activity	ATP + a protein = ADP + a phosphoprotein.
Enzyme regulation	Inhibited by staurosporine. Ref.3
Subunit structure	Monomer. Ref.1 Ref.3
Subcellular location	Cytoplasm Ref.1.
Sequence similarities	Belongs to the protein kinase superfamily. AFK protein kinase family. Contains 5 Kelch repeats.
Biophysicochemical properties	Kinetic parameters: KM=190 nM for actin-fragmin complex Ref.4 KM=25 µM for ATP Vmax=83 µmol/min/mg enzyme

Ontologies

Keywords
Cellular component	Cytoplasm
Domain	Kelch repeat Repeat
Ligand	ATP-binding Nucleotide-binding
Molecular function	Kinase Serine/threonine-protein kinase Transferase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Cellular component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW protein serine/threonine kinase activity 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

Chain

1 – 737

737

Actin-fragmin kinase

PRO_0000086044

Regions

Repeat

431 – 480

50

Kelch 1

Repeat

482 – 533

52

Kelch 2

Repeat

534 – 584

51

Kelch 3

Repeat

586 – 635

50

Kelch 4

Repeat

646 – 693

48

Kelch 5

Nucleotide binding

178 – 185

8

ATP Potential

Compositional bias

350 – 365

16

Poly-Ser

Sites

Binding site

82

1

ATP; via amide nitrogen

Binding site

98

1

ATP

Binding site

163

1

ATP; via carbonyl oxygen

Binding site

165

1

ATP; via amide nitrogen

Binding site

232

1

ATP

Experimental info

Sequence conflict

506

1

S → G AA sequence Ref.2

Sequence conflict

507

1

T → P AA sequence Ref.2

Sequence conflict

560

1

Q → K AA sequence Ref.1

Sequence conflict

560

1

Q → K AA sequence Ref.2

Sequence conflict

639

1

S → W AA sequence Ref.2

Secondary structure

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737

Helix Strand Turn

Details...

Helix

5 – 15

11

Helix

25 – 27

3

Turn

52 – 54

3

Helix

69 – 71

3

Beta strand

86 – 88

3

Beta strand

95 – 98

4

Helix

103 – 115

13

Beta strand

123 – 130

8

Helix

135 – 143

9

Beta strand

145 – 147

3

Helix

149 – 153

5

Beta strand

157 – 163

7

Beta strand

175 – 177

3

Helix

178 – 181

4

Helix

184 – 189

6

Helix

193 – 200

8

Beta strand

204 – 206

3

Beta strand

208 – 210

3

Helix

216 – 218

3

Beta strand

219 – 221

3

Beta strand

228 – 230

3

Helix

247 – 257

11

Helix

264 – 276

13

Helix

287 – 290

4

Helix

294 – 302

9

Helix

305 – 322

18

Helix

331 – 341

11

Sequences

	Sequence		Length	Mass (Da)
	P80197 [UniParc]. Last modified May 5, 2009. Version 2. Checksum: D950BD5BAE6764B0	FASTA	737	80,955
	10 20 30 40 50 60 MAGALWEIEK ELFTKLPAPS SAINSHLQPA KPKVPQKKPS KWDPPAEFKV DLSTAVSYND 70 80 90 100 110 120 IGDINWKNLQ QFKGIERSEK GTEGLFFVET ESGVFIVKRS TNIESETFCS LLCMRLGLHA 130 140 150 160 170 180 PKVRVVSSNS EEGTNMLECL AAIDKSFRVI TTLANQANIL LMELVRGITL NKLTTTSAPE 190 200 210 220 230 240 VLTKSTMQQL GSLMALDVIV NNSDRLPIAW TNEGNLDNIM LSERGATVVP IDSKIIPLDA 250 260 270 280 290 300 SHPHGERVRE LLRTLIAHPG HESSQFHSIR DIITLYTGYD VGTEGSISMQ EGFLATVREC 310 320 330 340 350 360 ASFDLDAFER ELLSWQESLQ KCHNLSISPQ AIPFILRMLR IFHDAIHNPS PSSPSPSPSS 370 380 390 400 410 420 SSSTSHPTPA SSSTSSTLPS SIPSSSNTSP PPASSSESLV GVEECAWLKV VVPNEKPAPR 430 440 450 460 470 480 RYHSGVLYEG KLYVFGGVCI KTASNDFYVF DFAKKKWSIV VAQGEAPSPR CGHSATVYGG 490 500 510 520 530 540 KMWIFGGHNN NKQPYSDLYT FDFAKSTWEK IEPTKDGPWP SPRYHHSATL VGASLYIFGG 550 560 570 580 590 600 AEHKSKYHND VYVYKFDANQ WELLNATGET PEPRAGQMTV EWNNSLFTFG GHGGEGGYTS 610 620 630 640 650 660 FVDAHVFEIA TNTFHEVDCS GTFPRTARPL SYVPYYYGSG DKREGAVFSF GGSDGKSPLG 670 680 690 700 710 720 SLYQWNLKTH KWKIIKAWMA VEDNTIGSMA AIASGKLDPI PRYGHCTVLD DTGVISIFGG 730 SGSLFLDDIV QFDMTES

References

[1]	"A novel type of protein kinase phosphorylates actin in the actin-fragmin complex." Eichinger L., Bomblies L., Vandekerckhove J., Schleicher M., Gettemans J. EMBO J. 15:5547-5556(1996) [PubMed: 8896448] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 42-49; 225-234; 410-420; 442-454; 471-481; 493-507; 545-574; 629-642; 644-656 AND 659-668, FUNCTION, SUBUNIT, ATP-BINDING SITE, SUBCELLULAR LOCATION.
[2]	"Purification and partial amino acid sequence of the actin-fragmin kinase from Physarum polycephalum." Gettemans J., de Ville Y., Vandekerckhove J., Waelkens E. Eur. J. Biochem. 214:111-119(1993) [PubMed: 8389700] [Abstract] Cited for: PROTEIN SEQUENCE OF 42-49; 225-234; 410-420; 442-454; 471-481; 493-507; 545-574; 629-642; 644-656 AND 659-668.
[3]	"Physarum actin is phosphorylated as the actin-fragmin complex at residues Thr203 and Thr202 by a specific 80 kDa kinase." Gettemans J., de Ville Y., Vandekerckhove J., Waelkens E. EMBO J. 11:3185-3191(1992) [PubMed: 1324166] [Abstract] Cited for: SUBUNIT, ENZYME REGULATION.
[4]	"Actin kinase: a protein kinase that phosphorylates actin of fragmin-actin complex." Furuhashi K., Hatano S. J. Biochem. 111:366-370(1992) [PubMed: 1587799] [Abstract] Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES.
[5]	"Phosphorylation by actin kinase of the pointed end domain on the actin molecule." Furuhashi K., Hatano S., Ando S., Nishizawa K., Inagaki M. J. Biol. Chem. 267:9326-9330(1992) [PubMed: 1315751] [Abstract] Cited for: FUNCTION.
[6]	"The crystal structure of the Physarum polycephalum actin-fragmin kinase: an atypical protein kinase with a specialized substrate-binding domain." Steinbacher S., Hof P., Eichinger L., Schleicher M., Gettemans J., Vandekerckhove J., Huber R., Benz J. EMBO J. 18:2923-2929(1999) [PubMed: 10357805] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.9 ANGSTROMS) OF 2-343 IN COMPLEX WITH AMP.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

U64722 mRNA. Translation: AAB08728.1.

PIR

S36952.
S72442.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1CJA	X-ray	2.90	A/B	2-343	[»]

ProteinModelPortal

P80197.

SMR

P80197. Positions 2-343.

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Family and domain databases

InterPro

IPR015275. Actin-fragmin_kin_cat.
IPR015915. Kelch-typ_b-propeller.
IPR006652. Kelch_1.
IPR011498. Kelch_2.
IPR011009. Kinase-like_dom.
IPR000403. PI3/4_kinase_cat.
[Graphical view]

Gene3D

G3DSA:2.120.10.80. Kelch-typ_b-propeller. 1 hit.
G3DSA:1.10.1070.11. PI3/4_kinase_cat. 1 hit.

Pfam

PF09192. Act-Frag_cataly. 1 hit.
PF01344. Kelch_1. 2 hits.
PF07646. Kelch_2. 1 hit.
[Graphical view]

SUPFAM

SSF56112. Kinase_like. 1 hit.

ProtoNet

Search...

Entry information

Entry name

AFK_PHYPO

Accession

Primary (citable) accession number: P80197
Secondary accession number(s): Q94706

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 1, 1993
Last sequence update:	May 5, 2009
Last modified:	January 25, 2012
This is version 63 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)