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cRaf-1
cRaf-1 is a protein serine/threonine kinase which plays a central role in the mitogenic response of cells to growth factors,
cytokines, and many oncogenes. The kinase has been implicated as a pivotal regulator of endothelial cell survival during
angiogenesis. It may also have a role in antineoplastic drug resistance. Inhibition of phosphorylation of Raf-1 by Akt
sensitizes human ovarian cancer cells to paclitaxel, a natural product which has significant anti-tumor activity particularly
in advanced ovarian and breast carcinomas.
Raf-1is activated through both Ras-dependent and Ras-independent mechanisms. Once activated, Raf-1 phosphorylates
and activates the dual specificity protein kinases MEK1 and MEK2 which in turn phosphorylate and activate the
serine/threonine specific MAP kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors and play an important
role in the control of diverse cellular processes.
The protein kinase activity of c-Raf-1 is strongly influenced by two key tyrosine residues upstream of the ATP binding site
(Y340 and Y341). Dephosphorylation of Ser259 by protein phosphatase 2A (GLO130-GLO132) contributes to Raf-1
activation by supporting its membrane accumulation rather than by increasing the specific activity of the kinase.
Phosphorylation of Raf-1 by PAK1 (GLO100-001, GLO100-005) and Src regulates Raf-1 autoinhibition. PAK1 primes MEK1
for phosphorylation by Raf-1 kinase during cross-cascade activation of the ERK pathway.
Catalog
GLO115-100
Price/vial
$378.00
Source
GST-tagged recombinant human produced in E. coli
Purity
> 80% by SDS-PAGE.
Supplied
100 ng in 50 ml 50 mM Tris-HCl pH 7.0 buffer containing 14 mM b-mercaptoethanol, 1 mM benzamidine,
0.1 mM phenylmethanesulfonyl fluoride, 1 mM EDTA and 50% glycerol.
Activity
20 units/mg with MEK as substrate. One unit of c-raf-1 is the amount of the enzyme that generates one
unit of MAP kinase per minute. One unit of MAP kinase is the amount that incorporates 1 nmol of
phosphate into myelin basic protein (MBP, GLO126-010, GLO126-025) per min at 30 EC, pH 7.0.
Storage
Maintain preparations in aliquots at -70 EC. Avoid repeated thawing.
References:
Bonner TI et al (1985) “Structure and biological activity of human homologs of the raf/mil oncogene” Mol Cell Biol
5, 1400; Stokoe D et al (1994) “Activation of Raf as a result of recruitment to the plasma membrane” Science 264,
1463; Alessi DR et al (1995) “Assay and expression of mitogen-activated protein kinase, MAP kinase kinase, and
Raf” Methods Enzymol 255, 2792; Bosch E et al (1997) Mutations of critical amino acids affect the biological and
biochemical properties of oncogenic A-Raf and Raf-1" Oncogene 15, 1021; Coles LC et al (2002) “PAK1 primes
MEK1 for phosphorylation by Raf-1 kinase during cross-cascade activation of the ERK pathway” Oncogene 21, 2236;
Kubicek M (2002) “Dephosphorylation of Ser-259 regulates Raf-1 membrane association” J Biol Chem 277, 7913;
Lee M et al (2003) “Down-regulation of Raf-1 kinase is associated with paclitaxel resistance in human breast cancer
MCF-7/Adr cells” Cancer Lett 193, 57; Mabuchi S et al (2002) “Inhibition of phosphorylation of BAD and Raf-1 by
Akt sensitizes human ovarian cancer cells to paclitaxel” J Biol Chem 277, 33490; Tran NH & Frost JA (2003)
“Phosphorylation of Raf-1 by p21-activated kinase 1 and Src regulates Raf-1 autoinhibition” J Biol Chem 278, 11221
GloboZymes products are for basic science research purposes only. They are not intended for human
drug, food additive, clinical or household use.