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Kinase Substrates

The protein kinase substrates below are supplied per vial as indicated in 50 ml 50 mM Tris-HCl, pH 7.0 buffer containing 14 mM b-mercaptoethanol, 1 mM benzamidine, 0.1 mM PMSF, 1 mM EDTA, 10% glycerol. Maintain in aliquots at -70°C. Avoid repeated thawing. For ordering information, click here.
eIF-4E; Protein Synthesis Initiation Factor 4E
GLO118 (1 mg, $285)

Cap-binding protein involved in rate limiting step in protein synthesis. Undergoes phosphorylation at Ser209 in response to growth factors and other extracellular stimuli. Occurs as a complex with eIF-4BP’s and as part of the eIF-4F complex. Source human recombinant. Purity > 90% by SDS-PAGE. Apparent Mr ~ 28 kDa. For eIF-4E phosphorylation site mutant preparations see initiation factors. For eIF-4E antibody, see antibodies.

ref: J Biol Chem 270: 14824

PDC; Pyruvate Dehydrogenase Complex
GLO124 (10 mg, $285)

Mitochondrial enzyme regulated by reversible phosphorylation. Catalyzes the oxidative decarboxylation of pyruvate. Purified preparations undergo phosphorylation because endogenous kinase is tightly associated with the complex. Source bovine heart. Purity > 90% by SDS-PAGE. Specific activity ~ 12-18 mmol of NADH formed per min per mg of protein with pyruvate as substrate. Apparent Mr ~ 8,500,000 by sedimentation equilibrium centrifugation. Subunit apparent Mr's ~ 74,000 (E2), 53,000 (E3), 41,000 (E1a), 48,000 (E3-BP) and 36,000 (E1b).

ref: 1. Methods Enzymol 166: 321 2. Methods Enzymol 89:376

BCKDH; Branched-Chain a-Keto Acid Dehydrogenase Complex
GLO125 (10 mg, $285)

Mitochondial enzymes regulated by reversible phosphorylation. Catalyzes the oxidative decarboxylation of the branched-chain a-keto acids derived from the essential amino acids, leucine, isoleucine and valine. Purified preparations undergo phosphorylation because endogenous kinase is tightly associated with the complex. Source bovine kidney. Purity > 90% by SDS-PAGE. Specific activity ~ 8-10 mmol of NADH formed per min per mg of protein with a-ketoisovalerate as substrate. Apparent Mr ~ 3,200,000 by sedimentation equilibrium centrifugation. Subunit apparent Mr's 52,000 (E2), 46,000 (E1a), 35,000 (E1b). NEW! BCKDH kinase kit: see GLO125-01 protein kinases.

ref: 1. Methods Enzymol 166: 321 2. Methods Enzymol 166: 309 3. Proc Natl Acad Sci USA 81: 4335 4. J Biol Chem 262: 5129

MBP; Myelin Basic Protein
GLO126 (1 mg, $285)

Useful as substrate for many protein kinases and protein phosphatases. Source bovine brain. Purity > 90% by SDS-PAGE. Apparent Mr ~ 20-kDa.

ref: J Biol Chem 258: 930

c-Jun; Transactivating Factor
GLO127 (1 mg, $285)

Important proto-oncogene and transcription factor. Source recombinant human. Purity > 90% by SDS-PAGE. Apparent Mr ~ 39-kDa.

ref: 1. Al-Murrani SWK, Damuni Z in press 2. Biochem J 341: 293
 

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