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Signal Transduction Inhibitors in Cellular Function

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Signal transduction pathways mediate cell-cell interactions and integrate signals from the extracellular environment through specific receptors at the cell membrane. They play a pivotal role in regulating cellular growth and differentiation and in mediating many physiological and pathological processes, such as apoptosis, inflammation, and tumor development. The mitogenactivated protein kinases (MAPKs) constitute a cascade of phosphorylation events that transmit extracellular growth signals through membrane-bound Ras to the nucleus of the cell. In this chapter, detailed protocols for analyzing the kinase activities of the key components of the MAPKs pathway—MEK1, ERK1, JNK, and p38 MAPK—are described. A brief introduction to the chemical inhibitors to the MAPKs pathway is provided in the method section of each kinase assay. Inhibitors of other signaling pathways are summarized in Table 1 . The reporter assay of cyclin D1, a key downstream target gene of MAPKs pathway, is also described in detail.
Table 1.  Signal Transduction Pathways and Inhibitors

Signal Transduction Pathways

Inhibitors

References

Tyrosine Kinase Growth Factor Receptors

   

HER2/Neu inhibitor

Herceptin (anti-HER2 antibody)

(53,54)

EGF receptors

IMC-C225 (anti-EGFR antidody)

(55,56)

EGFR-Tyrosine kinase

ZD1839, pyridopyrimidines

(54,57,58)

Ras Signaling

   

Inhibitors of Ras farnesyltransferase FPP analogues

   

CAAX peptide analogues

BZA-5B, L-739, Cys-4-ABA-Met FT1-276, SCH44342, SCH66336

(5759)

Bisubstrate inhibitors

BMS-182878, BMS-184467

(5759)

Inhibitors of the Raf protein kinases

ISIS5132, BAY43-9006

(5759)

Mitogen-Activated Protein Kinase Pathways

   

Inhibitors of MEK

PD184352, PD098059, U0126, R009-22110

(17,20,21 , 2327)

Inhibitors of ERK1 and ERK2

PD098059, E64D, calpeptin

(5,28)

Inhibitors of JNKs

SB600125

(33,34)

Inhibitors of p38 kinases

SB203580, SB202190, SB242235, SB239063, SB220025, SB202474, SC68376, FR167653

(36,37 , 3944,46)

Inhibitors of PI3-Kinase Signaling Pathways

Wortmannin, LY294002

(60,61)

Protein Phosphatases Inhibitors Proteasome Inhibitors

Microcystines, calyculins, cantharidin

(67)

Peptide aldehydes

ALLN, MG132, PSI, MG115

(63)

Peptide boronates

MG262

(63)

Nonpeptide inhibitors

Lactacystin

(63)

DCI

3,4-DCI

(63,64)

Peptide vinyl sulfones

NLVS, YL3YS

(63)

Epoxyketones

Epoxomicin, eponemycin, Ac-hFLFL-epoxide

(63,65)

Bivalent inhibitors

Polyoxyethylene

(66,67)

Natural compound inhibitors

TMC-95A, gliotoxin, EGCG

(63,68)

Histone Deacetylase inhibitors

   

Short-chain fatty acids

Butyrates

(70,75)

Hydroxamic acids

Trichostatin A, oxamflatin

(73)

Cyclic peptides

Trapoxin A, FR901228, apicidin

 

Benzamides

MS-27275

(78,80)

SIR2 inhibitors

Nicotinamide, splitomicin

(78,79)

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