KinaseNET

Nomenclature

Short Name:

MLK1

Full Name:

Mitogen-activated protein kinase kinase kinase 9

Alias:

EC 2.7.11.25
M3K9
MAPKKK9
MLK-1
PRKE1

Classification

Type:

Protein-serine/threonine kinase

Group:

TKL

Family:

MLK

SubFamily:

MLK

Specific Links

Entrez-Gene Entry:	4293
Entrez-Protein Entry:	NP_149132
GeneCards Entry:	PRKE
KinBASE Entry:	MLK1
OMIM Entry:	600136
Pfam Entry:	P80192
PhosphoNET Entry:	P80192
Phosphosite Plus Entry:	804
ScanSite Entry:	P80192
Source Entry:	MAP3K9
UCSD-Nature Entry:	A001549
UniProt Entry:	P80192
Kinexus Products:	MLK1 Mixed-lineage protein-serine kinase 1; Mitogen-activated protein kinase kinase kinase 9 pan-specific antibody AB-NK278-1 Mixed-lineage protein-serine kinase 1 / Mitogen-activated protein kinase kinase kinase 9 (A1086-S1104, human) peptide - Powder PE-01BDX99, PE-01BDY95

General Links

Structure

Mol. Mass (Da):

121895

# Amino Acids:

1104

# mRNA Isoforms:

mRNA Isoforms:

123,373 Da (1118 AA; P80192-4); 121,895 Da (1104 AA; P80192)

4D Structure:

Homodimer

1D Structure:

Retrieve Gene Sequence
Retrieve Full Protein Sequence
Retrieve Catalytic Domain Sequence

3D Image (rendered using PV Viewer):

PDB ID

4UY9

Subfamily Alignment

Domain Distribution:

Start	End	Domain
52	116	SH3
144	403	TyrKc
282	323	Coiled-coil
329	349	Coiled-coil
430	464	Coiled-coil
470	490	Coiled-coil
430	451	Leucine-zipper
465	486	Leucine-zipper
144	408	Pkinase

Kinexus Products

Click on entries below for direct links to relevant products from Kinexus for this protein kinase.

hiddentext

○	Mixed-lineage protein-serine kinase 1; Mitogen-activated protein kinase kinase kinase 9 pan-specific antibody AB-NK278-1
○	Mixed-lineage protein-serine kinase 1 / Mitogen-activated protein kinase kinase kinase 9 (A1086-S1104, human) peptide - Powder PE-01BDX99, PE-01BDY95

Post-translation Modifications

For detailed information on phosphorylation of this kinase go to PhosphoNET

Acetylated:

K594, K595, K607.

Serine phosphorylated:

S308+, S519, S533, S548, S552, S616, S652, S695, S807, S809, S812, S917, S938+.

Threonine phosphorylated:

T304+, T312+, T452, T535, T565, T808, T915, T948.

Tyrosine phosphorylated:

Y110.

Distribution

Based on gene microarray analysis from the NCBI

Human Tissue Distribution

% Max Expression:

Mean Expression:

Number of Samples:

Standard Deviation:

100

857

29

1279
5

42

17

43
4

37

2

13
48

412

81

489
52

449

25

375
4

35

74

14
26

224

35

394
55

475

30

552
35

296

17

252
6

53

86

39
6

50

24

57
64

552

176

590
4

38

24

13
3

27

14

30
5

47

19

45
4

37

16

33
2

18

195

20
6

50

13

48
6

49

83

34
44

380

109

443
7

56

17

54
4

34

21

31
7

56

4

13
8

71

12

53
3

28

17

34
79

677

45

606
6

51

27

24
5

44

13

45
4

38

13

40
31

267

28

177
68

587

24

566
91

780

36

976
0.4

3

34

1
89

763

52

657
31

267

35

241

Evolution

Species Conservation

PhosphoNET % Identity:
PhosphoNET % Similarity:
Homologene %
Identity:

100

100

100
97.6

98.2

98
99.1

99.5

99
-

-

96
-

-

-
81.4

84.3

97
-

-

-
92.9

94.7

96
43

55.3

96
-

-

-
-

-

-
-

-

84
53.9

66.5

74
59

69.5

67
-

-

-
30.8

46.1

-
38.7

53.9

-
22.1

38.7

-
39.5

55.7

-
-

-

-
-

-

-
-

-

36
-

-

-
-

-

-
-

-

-

For a wider analysis go to PhosphoNET Evolution in PhosphoNET

Binding Proteins

Examples of known interacting proteins

hiddentext

No.	Name – UniProt ID
1	SH3RF1 - Q7Z6J0

Regulation

Activation:

Phosphorylation of Thr-304, Ser-308 and Thr-312 increases phosphotransferase activity. Homodimerization via the leucine zipper domains is required for autophosphorylation and subsequent activation.

Inhibition:

Synthesis:

Degradation:

Known Upstream Kinases

For further details on these substrates click on the Substrate Short Name or UniProt ID. Phosphosite Location is hyperlinked to PhosphoNET predictions.

Based on in vitro and/or in vivo phosphorylation data

Kinase Short Name	UniProt ID (Human)	Phosphosite Location	Phosphosite Sequence	Effect of Phosphorylation

MLK1	P80192	T304	LAREWHRTTKMSAAG	+
MLK1	P80192	T312	TKMSAAGTYAWMAPE	+
MLK1	P80192	S519	LKDGNRISLPSDFQH

Known Downstream Substrates

For further details on these substrates click on the Substrate Short Name or UniProt ID. Phosphosite Location is hyperlinked to PhosphoNET predictions.

Based on in vitro and/or in vivo phosphorylation data

Substrate Short Name	UniProt ID (Human)	Phosphosite Location	Phosphosite Sequence	Effect of Phosphorylation

MLK1 (MAP3K9)	P80192	S519	LKDGNRISLPSDFQH
MLK1 (MAP3K9)	P80192	T304	LAREWHRTTKMSAAG	+
MLK1 (MAP3K9)	P80192	T312	TKMSAAGTYAWMAPE	+

Protein Kinase Specificity

Matrix of observed frequency (%) of amino acids in aligned protein substrate phosphosites

Matrix Type:

Predicted from the application of the Kinexus Kinase Substrate Predictor Version 2.0 algorithm, which was trained with over 10,000 kinase-protein substrate pairs and 8,000 kinase-peptide substrate pairs.

Domain #:

Inhibitors

For further details on these inhibitors click on the Compound Name and enter it into DrugKiNET or click on the ID's

Based on in vitro and/or in vivo phosphorylation data

Compound Name	KD, Ki or IC50 (nM)	PubChem ID	ChEMBL ID	PubMed ID

TTT-3002	IC50 < 2 nM
Lestaurtinib	Kd = 3.5 nM	126565		22037378
R406	Kd = 4.3 nM	11984591		22037378
Staurosporine	Kd = 10 nM	5279		18183025
CHEMBL1240703	Kd = 15 nM	52945601	1240703	19654408
N-Benzoylstaurosporine	Kd = 15 nM	56603681	608533	18183025
K-252a; Nocardiopsis sp.	IC50 = 22 nM	3813	281948	17266195
Cdk1/2 Inhibitor III	IC50 < 25 nM	5330812	261720	22037377
Gö6976	IC50 < 25 nM	3501	302449	22037377
NU6140	IC50 < 25 nM	10202471	1802728	22037377
CEP6331	IC50 = 26 nM	9823787	460990	18714982
Amgen TBK 1 inhibitor (Compound II)	IC50 < 40 nM
BX795	IC50 < 40 nM	10077147	577784
Aloisine A	IC50 > 50 nM	5326843	75680	22037377
Alsterpaullone	IC50 > 50 nM	5005498	50894	22037377
ALX-270-403-M001	IC50 > 50 nM			22037377
GSK-3 Inhibitor IX	IC50 > 50 nM	5287844	409450	22037377
IDR E804	IC50 > 50 nM	6419764	1802727	22037377
Pazopanib	IC50 > 50 nM	10113978	477772	22037377
PF-03814735	IC50 < 50 nM	49830590		20354118
SB218078	IC50 > 50 nM	447446	289422	22037377
KIN 112	IC50 < 60 nM
Wyeth PDK1 Inhibitor Compound 1	IC50 < 60 nM
MRT67307	IC50 < 80 nM	44464263
Purvalanol B	IC50 < 80 nM	448991	23254
AT9283	IC50 > 100 nM	24905142		19143567
BI-78D3	IC50 = 100 nM	2747117	508280
IPA-3	IC50 < 100 nM	521106	472940
PP242	IC50 = 100 nM	25243800
Purvalanol A	IC50 = 100 nM	456214	23327
Syk Inhibitor	IC50 = 100 nM	6419747	104279
CEP5104	IC50 = 103 nM	23378546	460989	18714982
K-252c	IC50 = 138 nM	3815	16958	17266195
Alsterpaullone; 2-Cyanoethyl	IC50 > 150 nM	16760286	260138	22037377
BCP9000906	IC50 > 150 nM	5494425	21156	22037377
Bosutinib	IC50 > 150 nM	5328940	288441	22037377
Compound 52	IC50 > 150 nM	2856		22037377
GSK-3 Inhibitor XIII	IC50 > 150 nM	6419766	359482	22037377
IKK-2 Inhibitor IV	IC50 > 150 nM	9903786	257167	22037377
Indirubin-3′-monoxime	IC50 > 150 nM	5326739		22037377
JAK3 Inhibitor VI	IC50 > 150 nM	16760524		22037377
TWS119	IC50 > 150 nM	9549289	405759	22037377
Nintedanib	Kd = 220 nM	9809715	502835	22037378
1;9-Pyrazoloanthrone	IC50 > 250 nM	8515	7064	22037377
Aloisine; RP106	IC50 > 250 nM	44350092	126343	22037377
Flt-3 Inhibitor II	IC50 > 250 nM	11601743	377193	22037377
PKCb Inhibitor	IC50 > 250 nM	6419755	366266	22037377
Semaxinib	IC50 > 250 nM	5329098	276711	22037377
Tozasertib	Kd = 390 nM	5494449	572878	18183025
BI-D1870	IC50 < 400 nM	25023738	573107
CZC-25146	IC50 < 400 nM
NVP-TAE684	Kd = 410 nM	16038120	509032	22037378
Bisindolylmaleimide I	IC50 = 500 nM	2396	7463	22037377
Bisindolylmaleimide IV	IC50 = 500 nM	2399	266487	22037377
CGP74514A	IC50 = 500 nM	2794188	367625	22037377
Dovitinib	IC50 = 500 nM	57336746		22037377
GSK-3 Inhibitor X	IC50 = 500 nM	6538818	430226	22037377
MK5108	IC50 > 500 nM	24748204		20053775
PKR Inhibitor	IC50 = 500 nM	6490494	235641	22037377
SB202190	IC50 = 500 nM	5353940	278041	22037377
SU11652	IC50 = 500 nM	24906267	13485	22037377
Syk Inhibitor II	IC50 = 500 nM	16760670		22037377
VEGFR2 Kinase Inhibitor II	IC50 = 500 nM	5329155	88606	22037377
GW441756 hydrochloride	IC50 < 600 nM	16219400
IKK-3 inhibitor IX	IC50 < 600 nM	11626927	373751
K00596a	IC50 < 600 nM	9549298	200027
Vemurafenib	IC50 < 600 nM	42611257	1229517
KW2449	Kd = 650 nM	11427553	1908397	22037378
Momelotinib	IC50 < 750 nM	25062766		19295546
AZ960	IC50 < 800 nM	25099184		18775810
Curcumin	IC50 < 800 nM	5281767	116438
GSK650394A	IC50 < 800 nM	25022668	558642
HG-10-102-01	IC50 < 800 nM
JNKIN7	IC50 < 800 nM	57340685
Ruxolitinib	IC50 < 800 nM	25126798	1789941
Alisertib	IC50 > 1 µM	24771867		22016509
AZ960	IC50 < 1 µM	25099184		18775810
BML-275	IC50 = 1 µM	11524144	478629
CHEMBL566515	IC50 = 1 µM	44478401	566515	19788238
Foretinib	Kd = 1 µM	42642645	1230609	22037378
Gö6983	IC50 > 1 µM	3499	261491	22037377
GSK461364A	IC50 = 1 µM	15983966	1908394
GW 843682X	IC50 = 1 µM	9826308	514499
HG-9-91-01	IC50 = 1 µM
IRAK-1-4 Inhibitor I	IC50 < 1 µM	11983295	379787
Kenpaullone	IC50 > 1 µM	3820	296586	22037377
PDK1/Akt/Flt Dual Pathway Inhibitor	IC50 > 1 µM	5113385	599894	22037377
Silmitasertib	IC50 > 1 µM	24748573		21174434
SNS314	IC50 > 1 µM	16047143	514582	18678489
Src Kinase Inhibitor I	IC50 > 1 µM	1474853	97771	22037377
Staurosporine aglycone	IC50 = 1 µM	3035817	281948
STO609	IC50 > 1 µM	51371511		22037377
SU9516	IC50 > 1 µM	5289419	258805	22037377
Syk Inhibitor IV	IC50 = 1 µM	10200390
VEGFR2 Kinase Inhibitor I	IC50 > 1 µM	6419834	86943	22037377
VEGFR2 Kinase Inhibitor IV	IC50 > 1 µM	5329468	92461	22037377
WZ3146	Kd > 1 µM	44607360		20033049
WZ4002	Kd > 1 µM	44607530		20033049
Ruboxistaurin	Kd = 1.4 µM	153999	91829	18183025
CHEMBL248757	Ki > 1.667 µM	44444843	248757	17935989
SU14813	Kd = 2.6 µM	10138259	1721885	18183025
Axitinib	Kd = 3 µM	6450551	1289926	22037378
IRAK-4 kinase inhibitor b	IC50 > 3 µM
TG101348	Kd = 3.2 µM	16722836	1287853	22037378
Sunitinib	Kd = 3.4 µM	5329102	535	18183025
JNJ-7706621	Kd = 3.6 µM	5330790	191003	18183025
Chelerythrine	IC50 < 4 µM	72311
JNJ-28871063	IC50 > 4 µM	17747413		17975007
Princeton's TrkA inhibitor compound 20h	IC50 < 4 µM
Ro-31-8220	IC50 < 4 µM	5083	6291
(5Z)-7-Oxozeaenol	IC50 > 4.5 µM		1077979
Akt Inhibitor VIII	IC50 > 4.5 µM	10196499	258844
GDC-0941	IC50 > 4.5 µM	17755052	521851
MSC 2032964A	IC50 > 4.5 µM

Disease Linkage

General Disease Association:

Cancer

Specific Cancer Types:

Non-small cell lung cancer (NSCLC)

Comments:

MLK1 may be an oncoprotein (OP). In non-small-cell lung cancers, a gain of function has been observed with MLK1 (MAP3K9), which results in downstream activation of ERK1 and ERK2 MAP kinases.

Gene Expression in Cancers:

TranscriptoNET (www.transcriptonet.ca) analysis with mRNA expression data retrieved from the National Center for Biotechnology Information's Gene Expression Omnibus (GEO) database, which was normalized against 60 abundantly and commonly found proteins, indicated altered expression for this protein kinase as shown here as the percent change from normal tissue controls (%CFC) as supported with the Student T-test in the following types of human cancers: Barrett's esophagus epithelial metaplasia (%CFC= -56, p<0.002); Cervical epithelial cancer (%CFC= +47, p<0.005); Oral squamous cell carcinomas (OSCC) (%CFC= -51, p<0.0001); Skin squamous cell carcinomas (%CFC= +83, p<0.055); and Vulvar intraepithelial neoplasia (%CFC= +112, p<0.018). The COSMIC website notes an up-regulated expression score for MLK1 in diverse human cancers of 408, which is 0.9-fold of the average score of 462 for the human protein kinases. The down-regulated expression score of 2 for this protein kinase in human cancers was 97% lower than the average score of 60 for the human protein kinases.

Mutagenesis Experiments:

Insertional mutagenesis studies in mice have not yet revealed a role for this protein kinase in mouse cancer oncogenesis. A K171A mutation can fully inhibit MLK1 phosphotransferase activity. Threonine phosphorylation can be lowered (through inactive kinase) with a T304A mutation, which also will reduce JNK activation. A mild effect on threonine phosphorylation and a minor impairment of JNK signalling can occur with a T305A mutation. A S308A mutation can reduce JNK signalling. A full loss of threonine phosphorylation and a strong inhibition of JNK signalling can occur with a T312A mutation.

Mutation Rate in All Cancers:

Percent mutation rates per 100 amino acids length in human cancers: 0.08 % in 24970 diverse cancer specimens. This rate is very similar (+ 1% higher) to the average rate of 0.075 % calculated for human protein kinases in general.

Mutation Rate in Specific Cancers:

Highest percent mutation rates per 100 amino acids length in human cancers: 0.57 % in 805 skin cancers tested; 0.36 % in 1093 large intestine cancers tested; 0.2 % in 589 stomach cancers tested; 0.16 % in 1807 lung cancers tested; 0.15 % in 602 endometrium cancers tested.

Frequency of Mutated Sites:

None > 4 in 20,222 cancer specimens

Comments:

Eight deletions (5 at E38delE), and no insertions or no complex mutations are noted on the COSMIC website.

COSMIC Entry:

MAP3K9

OMIM Entry:

600136

Nomenclature

Short Name:

Full Name:

Alias:

Classification

Type:

Group:

Family:

SubFamily:

Specific Links

General Links

Structure

Mol. Mass (Da):

# Amino Acids:

# mRNA Isoforms:

mRNA Isoforms:

4D Structure:

1D Structure:

3D Image (rendered using PV Viewer):

PDB ID

Subfamily Alignment

Domain Distribution:

Kinexus Products

hiddentext

Post-translation Modifications

Acetylated:

Serine phosphorylated:

Threonine phosphorylated:

Tyrosine phosphorylated:

Distribution

Human Tissue Distribution

% Max Expression: Mean Expression: Number of Samples: Standard Deviation:

% Max Expression: Mean Expression: Number of Samples: Standard Deviation:

Evolution

Species Conservation

PhosphoNET % Identity: PhosphoNET % Similarity: Homologene %Identity:

PhosphoNET % Identity: PhosphoNET % Similarity: Homologene %Identity:

Binding Proteins

hiddentext

Regulation

Activation:

Inhibition:

Synthesis:

Degradation:

Known Upstream Kinases

Based on in vitro and/or in vivo phosphorylation data

Known Downstream Substrates

Based on in vitro and/or in vivo phosphorylation data

Protein Kinase Specificity

Matrix Type:

Domain #:

Inhibitors

Based on in vitro and/or in vivo phosphorylation data

Disease Linkage

General Disease Association:

Specific Cancer Types:

Comments:

Gene Expression in Cancers:

Mutagenesis Experiments:

Mutation Rate in All Cancers:

Mutation Rate in Specific Cancers:

Frequency of Mutated Sites:

Comments:

COSMIC Entry:

OMIM Entry:

% Max Expression:

Mean Expression:

Number of Samples:

Standard Deviation:

% Max Expression:

Mean Expression:

Number of Samples:

Standard Deviation:

PhosphoNET % Identity:
PhosphoNET % Similarity:
Homologene %
Identity:

PhosphoNET % Identity:
PhosphoNET % Similarity:
Homologene %
Identity: