KINASE RECOGNITION SITE

• Submitted By: Mohit• M.Pharm(Pharmacology)

• Submitted to :Dr. Govind Singh (Associate Professor)• Dept. of Pharmaceutical

science,MDU,Rohtak

INTRODUCTION

• Protein Kinases are enzymes that modify the function of other proteins by attaching phosphate groups to them.

• 30% Human Protein Modified by Kinase.• Over 160 protein kinases are associated with

human diseases.• Involved in signal transduction pathway.

• Key controller of various biochemical pathways.

1) Glucose metabolism (glycolysis)2) Cell proliferation3) Transcription4) Cell migration.

The kinase Reaction

SIGNAL TRANSDUCTION PATHWAY

VARIOUS TYPE OF KINASE

1) PKA• Protein Kinase A/ cyclic AMP-dependent protein kinase

(cAPK), activated by cAMP production downstream of G protein coupled receptors.

2) PKC• Protein Kinase C. A large family responsive to lipid signaling. 3) PDK1• 'Master Kinase' that activates most other AGC kinases. A key

component in transducing signals from phosphoinsoitides to several other AGC-Group kinases like akt,pkc

4) PKN• The C-terminal kinase domain is very similar to PKC, but

these also have and N-terminal antiparallel coiled-coil structure (HR1 domain) that binds . This domain is also seen in the DMPK family which suggests a dual function for PKC1 (it also has the C1 and C2 domains typical of PKC).

5) AKT and SGK Akt (also known as Protein Kinase B, PKB) is downstream of

PDK1 in insulin and growth factor signaling.6) GRK• G-protein-coupled receptor kinases. As the name replies,

they modulate GPCRs, including the rhodopsin light-sensitive

• 7) DMPK Named after the Myotonic Dystrophy Protein

Kinase .it is an 80 kDa protein expressed exclusively in smooth,skeletal and cardiac muscles.

• 8) YANK• A family about which so little is known, it stands

for "Yet Another Novel Kinase". Animal-specific.

CLASSIFICATION OF KINASE

• Protein kinases have been classified according to their sequence and structure, which frequently, though not always, correlates with their biological functions.

• kinases are further subdivided into about 100 families and about 150 subfamilies

GROUP KINASE ACTIVITY1) PROTEIN S/T/Y KINASE (9799 sequences)

PROTEIN KINASEPHOSPHORYLASE KINASETYROSINE KINASEMYOSIN LIGHT CHAIN KINASECALCIUM-CALMOUDIN DEPENDENT PROTEIN KINASE

2) Rossmann-like (3777 sequences)

THYMIDINE KINASEADENYLYLSULFHATE KINASEPANTOTHENATE KINASE

3) ferredoxin-like foldkinases,( 1798 sequences)

NUCLEOSIDE –DIPHOSPHOKINASECREATINE KINASEARGININE KINASE

4) ribonuclease H-like (723 sequences)

HEXO KINASEGLYCEROL KINASE

5) TIM beta/alpha-barrelkinase,(231 sequences)

PYRUVATE KINASE

GROUPS KINASE ACTIVITY

6) GHMP kinase,( 382Sequences)

Mevalonate kinase Homoserine kinase

7) AIR synthetase-like, 251sequences

THIAMINE PHOSPHATE KINASE

8) RIBOFLAVIN KINASE 63 SEQUENCE RIBOFLAVIN KINASE

9) PHOSPHATE KINASE ,63 SEQUENCE PHOSPHATE KINASE

CROSS TALK OF KINASE

• It is the phenomenon by which channel of a transmission system create an undesired effect on another system.

• E.g : activation of one gene under the control of transcription factor and repression of another.

CROSS TALK MODELS

• cis crosstalk : it refers to communication between modifications on the same protein.

• ADJACENT CROSSTALK : It refers to residues that are close to one another in both the primary and tertiary structures.

• DISTAL CROSSTALK :It refers to residues that are far apart in both the primary and tertiary structures.

• Trans crosstalk : it occurs between modifications on two different proteins .

• Direct crosstalk :it refers to one PTM directly affecting the modification of a second residue (e.g., modification of one residue prevents the modification of another residue)

• Indirect crosstalk :It involves in modulation of protein-protein interaction via the presence, or lack, of a PTM (e.g., a PTM enhances the binding of a transcription factor leading to the recruitment of other coactivators)

Protein Kinase Substrate Recognition

• protein kinases for a number of different specificity motifs.

• Phosphoacceptor residue is indicated in red, amino acids which can function interchangeably at a particular residue are separated by a slash (/)

• residues which do not appear to contribute strongly to recognition are indicated by an "X".

Protein Kinase Recognition Determinant

cAMP-dependent Protein Kinase (PKA), Catalytic Subunit

R-R-X-S/TYY

Casein Kinase I (CK1) pS-X-X-S/T

Casein Kinase II (CK2) pS-X-X-S/T

Glycogen Synthase Kinase 3 (GSK-3)

S/T-X-X-X-pS-pT

CDK1-cyclin B S/T-P-X-R/K

Calmodulin-dependent Protein Kinase II (CaMK II)

R-X-X-S/T

p42 MAP Kinase (MAPK) S/T-P

• A = alanine, C = cysteine, D = aspartic acid, E = glutamic acid, F = phenylalanine, G = glycine, H = histidine, I = isoleucine, K = lysine, L = leucine, M = methionine, N = asparagine, P = proline, Q = glutamine, R = arginine, S = serine, T = threonine, W = tryptophan, V = valine, Y= tyrosine, X = any amino acid

VARIOUS YPE OF CROSSTALK

1) Arginine Methylation Blocks Phosphorylation

2) Phosphorylation Blocks Arginine Methylation

3) Distal Crosstalk Between Arginine Methylation and Phosphorylation

4) Lysine Methylation Blocks Phosphorylation

Arginine Methylation Blocks Phosphorylation

• cis crosstalk between PRMT1- and Akt.• The phosphorylation of other proteins that contain an

Akt consensus sequence (i.e., RXRXXS/T) are modulated by the methylation of adjacent arginine residues.

• Akt substrates (i.e., BAD, PGC-1α, eNOS, p27, GSK3β, and MDM2) were tested as PRMT1

substrates.• arginine methylation prevented phosphorylation of an

adjacent serine residue.• demonstrate a functional role for the inhibition of

serine phosphorylation by arginine methylation

Phosphorylation Blocks Arginine Methylation

• The Direct adjacent cis crosstalk, where phosphorylation of RNA polymerase II (RNAPII) prevents its methylation at R1810.

• The CTD (carboxy terminal domain) of RNAPII contains a series of YSPTSPS

• It contain arginine or lysine substitutions in the last position (i.e., YSPTSP[R/K])

• Phosphorylation of these residues activates RNAPII and aids in the recruitment of essential proteins (39, 40)

that are important for gene transcription (38). Due to the unique nature of the arginine and lysine substitutions.

• The functional consequence of a lack of methylation of R1810 is downregulated transcription of small nuclear RNA (snRNA) and small nucleolar RNA (snoRNA)

Distal Crosstalk Between Arginine Methylation and Phosphorylation

• distal crosstalk involves the members of the STAT (signal transducer and activator of transcription) family of proteins.

• which play important roles in cell• differentiation, survival, and apoptosis.• STAT (R31) methylated by PMRT.• STAT (R 27) PHOSPHORYLATED.• THAT shows changes in DNA binding ,decrease

protein stability.

Lysine Methylation Blocks Phosphorylation

• Direct adjacent cis crosstalk has been observed between lysine methylation and serine phosphorylation.

• E.g : pRb (retinoblastoma protein), a tumor suppressor protein that plays a critical role in controlling progression through the cell cycle.

• The methylation and phosphorylation on pRb that changes the transcriptional activity of DNA.