Phosphorylation of human fibrinogen in vitro with calcium‐activated, phospholipid‐dependent protein kinase and [<sup>32</sup>P]ATP

Papanikolaou, Paraskevi; Humble, Elisabet; Engström, Lorentz

doi:10.1016/0014-5793(82)80098-7

Cited by 27 publications

(5 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All other chemicals were of at least analytical grade. Lipid vesicles were prepared as described by Papanikolau et al [13].…”

Section: Methodsmentioning

confidence: 99%

A potent and highly selective peptide substrate for protein kinase C assay

Toomik

1997

Biochemical Journal

View full text Add to dashboard Cite

Protein kinases exhibit substrate specificities that are often primarily determined by the amino acids around the phosphorylation sites. Peptides corresponding to protein kinase C phosphorylation sites in several different proteins were synthesized on SPOTs membrane which has recently been found to be applicable for studies of protein kinase specificity. After phosphorylation with protein kinase C, we chose the best phosphorylated peptides for the investigation of the importance of amino acids immediately adjacent to the phosphorylation site. The selectivity of the best protein kinase C substrates from this study was analysed with protein kinases A, CK1 and CK2. According to these tests, the most favourable characteristics of SPOTs-membrane-associated peptides were demonstrated by peptide KRAKRKTAKKR. Kinetic analysis of peptide phosphorylation with protein kinase C revealed an apparent Km of 0.49 +/- 0.13 microM and Vmax of 10.0 +/- 0.5 nmol/min per mg with soluble peptide KRAKRKTAKKR. In addition, we assayed several other soluble peptides commonly used as protein kinase C substrates. Peptide KRAKRKTAKKR showed the lowest Km and the highest Vmax/Km value in comparison with peptides FKKSFKL, pEKRPSQRSKYL and KRAKRKTTKKR. Furthermore, of the peptides tested, KRAKRKTAKKR was the most selective substrate for protein kinase C. The favourable kinetic parameters combined with the selectivity should make the KRAKRKTAKKR peptide useful as a substrate for protein kinase C in the assays of both purified enzyme and in crude cell extracts.

show abstract

“…All other chemicals were of at least analytical grade. Lipid vesicles were prepared as described by Papanikolau et al [13].…”

Section: Methodsmentioning

confidence: 99%

A potent and highly selective peptide substrate for protein kinase C assay

Toomik

1997

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…Several protein kinases have been shown to phosphorylate fibrinogen in vitro. These include the cyclic AMP-dependent protein kinase from pig muscle (Engstrom et al, 1980;Humble et al, 1981), the Ca2+-activated phospholipid-dependent protein kinase from rat brain (Papanikolau et al, 1982) and pig spleen (Humble et al, 1984), a human platelet kinase activity enhanced by interferon (Galabru et al, 1983;Krust et al, 1983) and rat liver cytosol casein kinase 1 (Itarte et al, 1983), but no fibrinogen kinase activity seems to be present in human blood plasma (Humble etal., 1981).…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of fibrinogen by casein kinase 2

Guasch¹,

Plana²,

Pena³

et al. 1986

Biochemical Journal

View full text Add to dashboard Cite

Casein kinase 2 from rat liver cytosol phosphorylated human fibrinogen in a reaction that was not stimulated by Ca2+ or cyclic AMP, but was markedly inhibited by heparin, and proceeded at a similar rate when either ATP or GTP was used as phosphate donor. Analysis of casein kinase 2 by glycerol-density-gradient centrifugation showed that the activities towards fibrinogen, casein, phosvitin, high-mobility-group protein 14 and glycogen synthase coincided. Maximal incorporation into fibrinogen by casein kinase 2 averaged 1 mol of phosphate/mol of protein substrate, most of it in the alpha-chain, although some phosphorylation of the beta-chain was also detected. Analysis of phosphorylated alpha-chain revealed that most of the phosphate was incorporated on serine. Phosphorylation of human fibrinogen was also performed by casein kinase 2 from human polymorphonuclear leucocytes, lymphocytes and platelets.

show abstract

“…We conclude that B-50 protein kinase is a phospholipid-sensitive protein kinase that is very similar to kinase C. In this regard it must be noted that Papanikolaou et al (1982) have also reported a phospholipid-sensitive protein kinase which uses fibrinogen as a substrate. This enzyme was also shown to be very similar to kinase C (Papanikolaou et al, 1982). It may be that there exist many phospholipid- Vol.…”

Section: Figmentioning

confidence: 54%

Phosphorylation of B‐50 Protein by Calcium‐Activated, Phospholipid‐Dependent Protein Kinase and B‐50 Protein Kinase

Aloyo¹,

Zwiers²,

Gispen³

1983

Journal of Neurochemistry

224

View full text Add to dashboard Cite

B-50 is a brain-specific phosphoprotein, the phosphorylation state of which may play a role in the regulation of (poly)phosphoinositide metabolism. Several kinases were tested for their ability to phosphorylate purified B-50 protein. Only calcium-activated, phospholipid-dependent protein kinase (kinase C) and B-50 protein kinase were able to use B-50 protein as a substrate. Furthermore, kinase C specifically phosphorylates B-50 when added to synaptic plasma membranes. We further characterized the sensitivity of kinase C and B-50 kinase to ACTH (and various fragments), phospholipids, chlorpromazine, and proteolytic activation. Since the sensitivities of both kinases were similar, we conclude that B-50 protein kinase is a calcium-dependent, phospholipid-stimulated protein kinase of the same type as kinase C.

show abstract

Phosphorylation of human fibrinogen in vitro with calcium‐activated, phospholipid‐dependent protein kinase and [³²P]ATP

Cited by 27 publications

References 15 publications

A potent and highly selective peptide substrate for protein kinase C assay

A potent and highly selective peptide substrate for protein kinase C assay

Phosphorylation of fibrinogen by casein kinase 2

Phosphorylation of B‐50 Protein by Calcium‐Activated, Phospholipid‐Dependent Protein Kinase and B‐50 Protein Kinase

Contact Info

Product

Resources

About

Phosphorylation of human fibrinogen in vitro with calcium‐activated, phospholipid‐dependent protein kinase and [32P]ATP

Cited by 27 publications

References 15 publications

A potent and highly selective peptide substrate for protein kinase C assay

A potent and highly selective peptide substrate for protein kinase C assay

Phosphorylation of fibrinogen by casein kinase 2

Phosphorylation of B‐50 Protein by Calcium‐Activated, Phospholipid‐Dependent Protein Kinase and B‐50 Protein Kinase

Contact Info

Product

Resources

About

Phosphorylation of human fibrinogen in vitro with calcium‐activated, phospholipid‐dependent protein kinase and [³²P]ATP