[28] Use and specificity of staurosporine, UCN-O1, and calphostin C as protein kinase inhibitors

Tamaoki, Tatsuya

doi:10.1016/0076-6879(91)01030-6

Cited by 330 publications

(172 citation statements)

References 17 publications

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“…rhZAP70 327-606 yielded diffractionquality crystals in the presence of staurosporine and AMP-PCP (see "Experimental Procedures"). Staurosporine, a microbial alkaloid from Streptomyces, is a nonselective, ATP-competitive inhibitor of many kinases (23)(24)(25)(26)(27). In the TR-FRET assay, staurosporine inhibited rhZAP70 327-606 with an IC 50 of 55.8 nM (Fig.…”

Section: Resultsmentioning

confidence: 99%

The Three-dimensional Structure of the ZAP-70 Kinase Domain in Complex with Staurosporine

Jin

Pluskey²,

Petrella

et al. 2004

Journal of Biological Chemistry

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The ZAP-70 tyrosine kinase plays a critical role in T cell activation and the immune response and therefore is a logical target for immunomodulatory therapies. Although the crystal structure of the tandem Src homology-2 domains of human ZAP-70 in complex with a peptide derived from the subunit of the T cell receptor has been reported (Hatada, M. H., Lu, X., Laird, E. R., Green, J., Morgenstern, J. P., Lou, M., Marr, C. S., Phillips, T. B., Ram, M. K., Theriault, K., Zoller, M. J., and Karas, J. L. (1995) Nature 377, 32-38), the structure of the kinase domain has been elusive to date. We crystallized and determined the three-dimensional structure of the catalytic subunit of ZAP-70 as a complex with staurosporine to 2.3 Å resolution, utilizing an active kinase domain containing residues 327-606 identified by systematic N-and C-terminal truncations. The crystal structure shows that this ZAP-70 kinase domain is in an active-like conformation despite the lack of tyrosine phosphorylation in the activation loop. The unique features of the ATP-binding site, identified by structural and sequence comparison with other kinases, will be useful in the design of ZAP-70-selective inhibitors.The zeta-associated protein, 70 kDa (ZAP-70), 1 a Syk family tyrosine kinase associated with the subunit of the T cell receptor, is primarily expressed in T and NK cells and plays an essential role in signaling through the T cell receptor (TCR) (2, 3). TCR-mediated activation of T cells is crucial to the immune response. Transplant rejection and diseases, such as allergic responses and autoimmune disorders, stem from a failure to adequately modulate T cell activation. In humans, ZAP-70 gene mutations have been identified that confer lower ZAP-70 protein expression levels or catalytically inactive ZAP-70 proteins (4 -6). ZAP-70 deficiency results in the absence of mature CD8 ϩ T cells and the prevention of TCR-mediated activation of CD4 ϩ T cells, and it can lead to severe combined immunodeficiency. Peptides that block the association of ZAP-70 with the subunit (7,8) and peptides that antagonize ZAP-70 tyrosine kinase activity (9) block T cell activation in vitro. These studies indicate that ZAP-70 antagonists could be useful immunomodulatory therapeutic agents.ZAP-70 contains two N-terminal SH2 (Src homology domain 2) domains and a C-terminal kinase domain. During T cell activation, the binding of ZAP-70 SH2 domains to the phosphorylated subunit on the activated TCR complex causes a colocalization with the Lck tyrosine kinase that phosphorylates ZAP-70 on Tyr 493 in the activation loop (10, 11). Also, ZAP-70 autophosphorylates multiple tyrosines in the region between the SH2 domains and the kinase domain (10, 12), including the binding sites for additional SH2-containing signaling proteins such as SLP-76, Lat, Lck, PLC␥1, Vav, Shc, Ras-GAP, and Abl (reviewed in Refs. 8, 13, and 15). ZAP-70-mediated activation of these downstream effectors leads to the release of intracellular calcium stores, and the transcription of interleukin-2 and other ...

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Section: Resultsmentioning

confidence: 99%

The Three-dimensional Structure of the ZAP-70 Kinase Domain in Complex with Staurosporine

Jin

Pluskey²,

Petrella

et al. 2004

Journal of Biological Chemistry

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“…In addition, the selective PKC inhibitors calphostin C [41] and rottlerin [40], which are structurally distinct from staurosporine, induced considerable L-selectin shedding at concentrations shown to inhibit PKC in whole-cell assays [42][43][44][45]. Calphostin C, known to inhibit PKC by interacting with its diacyglycerol/ phorbol ester binding (regulatory) region [56], did not induce the up-regulation of Mac-1 expression, and blocked Mac-1 up-regulation upon neutrophil stimulation with PMA. These results indicate that calphostin C did not overtly activate neutrophils, unlike staurosporine, and effectively inhibited PKC.…”

Section: Discussionmentioning

confidence: 99%

Effects of selective protein kinase C inhibitors on the proteolytic down-regulation of L-selectin from chemoattractant-activated neutrophils

Alexander

Kishimoto

Walcheck

2000

Journal of Leukocyte Biology

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The signaling factors that direct the rapid shedding of L-selectin from neutrophils upon chemoattractant stimulation are poorly understood. Protein kinase C (PKC) has been implicated, yet previous studies have relied on the use of phorbol esters and nonselective kinase inhibitors. We treated neutrophils with various selective kinase inhibitors to evaluate their effects on N-formylmethionyl-leucyl-phenylalanine (fMLP)-induced Lselectin shedding. We found that three selective inhibitors of PKC, structurally related to staurosporine, largely blocked both fMLP-and phorbol 12-myristate 13-acetate (PMA)-induced L-selectin shedding; however, these inhibitors did not affect fMLP-induced up-regulation of Mac-1 (CD11b/ CD18) expression, which has been shown not to involve PKC. Other selective serine, threonine, and tyrosine kinase inhibitors were found not to block fMLP-induced L-selectin shedding. These findings provide more definitive evidence for the role of PKC in chemoattractant-induced L-selectin proteolysis. It is interesting that certain highly selective PKC inhibitors, not structurally related to staurosporine, were found to directly induce Lselectin shedding from neutrophils. J. Leukoc. Biol. 67: 415-422; 2000.

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“…Staurosporine is a potent nonspecific protein and tyrosine kinase inhibitor, with a very low therapeutic index in animals (Tamaoki, 1991). Thus, efforts to find staurosporine analogues of staurosporine have identified compounds specific for protein kinases.…”

Section: Mechanism Of Antiproliferative Activitymentioning

confidence: 99%

Small-molecule cyclin-dependent kinase modulators

2003

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[28] Use and specificity of staurosporine, UCN-O1, and calphostin C as protein kinase inhibitors

Cited by 330 publications

References 17 publications

The Three-dimensional Structure of the ZAP-70 Kinase Domain in Complex with Staurosporine

The Three-dimensional Structure of the ZAP-70 Kinase Domain in Complex with Staurosporine

Effects of selective protein kinase C inhibitors on the proteolytic down-regulation of L-selectin from chemoattractant-activated neutrophils

Small-molecule cyclin-dependent kinase modulators

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