Presentaci

1
The deep characterization of Aurora A
N-terminus
Isabel Peset Martin. Vanessa Ferreira Leire
Mendizabal Oiarzabal
2
Human Aurora kinases AC exhibit differential
substrate affinity, subcell localization and
associated activities
Paul D Andrews Oncogene (2005)
3
DOMAIN STRUCTURE OF Aurora kinases AC
Source Domain Start End PfamA PKinase
133 383 SMART STKc(hidden) 133 383
AURORA A
Source Domain Start
End PfamB Pfam-B 34136 1
62 SMART STKc(hidden)
77 327 PfamA PKinase
77 327 seg Low
Complexity (hidden) 204 215
AURORA B
Source Domain Start End PfamA PKinase
43 293 SMART STKc(hidden) 43 293
AURORA C
4
Amino acid sequence alignement of human Aurora
kinases AC.
aurB 1 MAQKENSYPWPYGRQTAPSG-------------------L
STLPQRVLR----KEPVTPSaurC 1 -------------------
---------------------MSSPRAVVQ----LGKAQPAaurA
1 MDRSKENCISGPVKATAPVGGPKRVLVTQQFPCQNPLPVNSGQAQRVL
CPSNSSQRVPLQaurB 38 ALVLMSRSN---------VQPTAAP
G------------------------QKVMENSSGTaurC 17
GEELATAN-----------QTAQQPS------------------------
SPAM------aurA 61 AQKLVSSHKPVQNQKQKQLQATSVPHPVS
RPLNNTQKSKQPLPSAPENNPEEELASKQKNaurB 65
PDILTRHFTIDDFEIGRPLGKGKFGNVYLAREKKSHFIVALKVLFKSQIE
KEGVEHQLRRaurC 36 -----RRLTVDDFEIGRPLGKGKFGNVYL
ARLKESHFIVALKVLFKSQIEKEGLEHQLRRaurA 121
EESKKRQWALEDFEIGRPLGKGKFGNVYLAREKQSKFILALKVLFKAQLE
KAGVEHQLRRaurB 125 EIEIQAHLHHPNILRLYNYFYDRRRIY
LILEYAPRGELYKELQKSCTFDEQRTATIMEELaurC 91
EIEIQAHLQHPNILRLYNYFHDARRVYLILEYAPRGELYKELQKSEKLDE
QRTATIIEELaurA 181 EVEIQSHLRHPNILRLYGYFHDATRVYLI
LEYAPLGTVYRELQKLSKFDEQRTATYITELaurB 185
ADALMYCHGKKVIHRDIKPENLLLGLKGELKIADFGWSVHAPSLRRKTMC
GTLDYLPPEMaurC 151 ADALTYCHDKKVIHRDIKPENLLLGFRGE
VKIADFGWSVHTPSLRRKTMCGTLDYLPPEMaurA 241
ANALSYCHSKRVIHRDIKPENLLLGSAGELKIADFGWSVHAPSSRRTTLC
GTLDYLPPEMaurB 245 IEGRMHNEKVDLWCIGVLCYELLVGNP
PFESASHNETYRRIVKVDLKFPASVPTGAQDLIaurC 211
IEGRTYDEKVDLWCIGVLCYELLVGYPPFESASHSETYRRILKVDVRFPL
SMPLGARDLIaurA 301 IEGRMHDEKVDLWSLGVLCYEFLVGKPPF
EANTYQETYKRISRVEFTFPDFVTEGARDLIaurB 305
SKLLRHNPSERLPLAQVSAHPWVRANSRRVLPPSALQSVA---aurC
271 SRLLRYQPLERLPLAQILKHPWVQAHSRRVLPPCAQMAS----au
rA 361 SRLLKHNPSQRPMLREVLEHPWITANSSKPSNCQNKESASKQ
S
5
Amino acid sequence alignment of Aurora A kinase
among species.
6
Post-translational modification prediction

• NetNGlyc - Prediction of N-glycosylation sites in
  human
• proteins

MDRSKENCISGPVKATAPVGGPKRVLVTQQFPCQNPLPVNSGQAQRVLCP
SNSSQRVPLQAQKLVSSHKPVQNQKQKQLQ 80
ATSVPHPVSRPLNNTQKSK QPLPSAPENNPEEELASKQKNEESKK
..................................................
.N............................ 80
............................................. 160
SeqName Position
Potential Jury
agreement result Sequence 52
NSSQ 0.5574
(5/9)
Sequence 93 NNTQ
0.4741 (4/9)
-
7
Post-translational modification prediction
SignalP - Prediction of signal peptide cleavage
sites Not found
HMMTOP - Prediction of transmembrane helices and
topology of proteins (Hungarian Academy of
Sciences) Not found
SUMOplot - Prediction of SUMO protein attachment
sites
HCA-Hydrophobic Cluster Analysis Not found
8
Post-translational modification prediction
NetPhos - Prediction of Ser, Thr and Tyr
phosphorylation sites in eukaryotic proteins
Phosphorylation sites predicted Ser 4 Thr 0
Tyr 0
Serine predictions Name Pos Context
Score Pred _________________________v__________
_______ Sequence 4 -MDRSKENC 0.045
. Sequence 10 ENCISGPVK 0.006
. Sequence 41 LPVNSGQAQ 0.125
. Sequence 51 VLCPSNSSQ 0.523
S Sequence 53 CPSNSSQRV 0.013
. Sequence 54 PSNSSQRVP 0.331
. Sequence 66 QKLVSSHKP 0.800
S Sequence 67 KLVSSHKPV 0.368
. Sequence 83 LQATSVPHP 0.056
. Sequence 89 PHPVSRPLN 0.007
. Sequence 98 NTQKSKQPL 0.323
. Sequence 104 QPLPSAPEN 0.634
S Sequence 116 EELASKQKN 0.352
. Sequence 123 KNEESKK-- 0.650 S
Threonine predictions Name Pos
Context Score Pred _________________________v_
________________ Sequence 16 PVKATAPVG
0.036 . Sequence 28 RVLVTQQFP 0.011
. Sequence 82 QLQATSVPH 0.087
. Sequence 95 PLNNTQKSK 0.498 .
9

• Scansite kinase site binding domain profiles

Motifs scanned AllStringency Medium
10
PDB code 1MQ4 TITLE CRYSTAL STRUCTURE OF
AURORA-A PROTEIN KINASE
11
CONCLUSION

• The Aurora Kinase A is very conserve throughout
  evolution.
• Human Auroras A-C have a conserved kinase domain
  and
• exhibit a relatively high sequence divergence at
  N terminal.
• SUMO protein attachment sites.
• Phosphorylation sites predicted (Ser 4)
• Possible binding domains SH3
• Aurora-A is crystalized

12
THANKS!!!