Assay of protein kinases with peptide substrates

Details Assay of protein kinases with peptide substrates Assay of protein kinases with peptide substrates

1988-12-12

1992-08-25

Rosen, Sam

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving transferase

435 21, 436161, 530327, 530329, 530344, 530352, C12Q 148, C07K 1524

Patent

active

051418525

ABSTRACT:
A method for assaying protein kinases that phosphorylate peptides such as Kemptide, such as cAMP-dependent protein kinase, or a glycogen synthase peptide, which is an excellent substrate for protein kinase C. Upon sequentially processing of reaction mixtures through tandem columns of cation and anion exchange resins improved separation of ATP from phosphorylated peptides is achieved such that radioactivity in background samples is nearly nil and the yield of phosphorylated peptides is high. This method is generally applicable to any protein kinase so long as the substrate peptide is appropriately structured such that the peptide retains a net positive charge when fully phosphorylated so that the peptide will adhere to the cation exchange resin and pass through the anion exchange resin. This method reduces labor, radioactivity, enzyme requirements, and costs of assaying protein kinases.

REFERENCES:
Sofer and Britton Biotechniques 1:198-203, 1983.
Kemp et al. Biochemistry 73:1038-1042 1976.
House et al. J. Biol. Chem. 262:772-777, 1987.
Glass et al. Anal. Biochem. 87:566-575, 1978.

Affiliated with

Also associated with

Rating

Assay of protein kinases with peptide substrates does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Assay of protein kinases with peptide substrates, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Assay of protein kinases with peptide substrates will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-384327