Phosphorylation by protein kinase C of a synthetic heptapeptide bearing a lysine residue on the C terminal side of serine

Kondo, Hiroki; Baba, Yoshimi; Takaki, Katsunori; Kondo, Kiyoshi; Kagamiyama, Hiroyuki

doi:10.1016/0006-291x(87)90464-5

Cited by 10 publications

(5 citation statements)

References 22 publications

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“…Until now there have been no reports of protein kinases other than PK-C which can phosphorylate the peptide A A A S F K A K K [41]. This peptide contains lysines as basic residues and phenylalanine on the C-terminal side of the serine A similar result was obtained from PK-C [20] using a peptide (ASGSFKL) homologous to the histone H 1 consensus (Table 1). The peptide sub- Table 1.…”

Section: Discussionsupporting

confidence: 62%

“…It was shown that the enzyme referred to as histone kinase II was identical to the proteolytically activated fragment of the Ca2÷/phospholipiddependent protein kinase [41]. However, it is worthwhile to mention that the nonapepiide synthetized by one of us and a very similar hexapeptide constructed later by Kondo et al [20] is considered as the most specific synthetic substrate for PK-C [4,20,[39][40][41].…”

Section: Introductionmentioning

confidence: 99%

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The phosphorylation site of Ca2+-dependent protein kinase from alfalfa

et al. 1989

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A 50 kDa, calcium-dependent protein kinase (CDPK) was purified about 1000-fold from cultured cells of alfalfa (Medicago varia) on the basis of its histone H1 phosphorylation activity. The major polypeptide from bovine histone H1 phosphorylated by either animal protein kinase C (PK-C) or by the alfalfa CDPK gave an identical phosphopeptide pattern. The phosphoamino acid determination showed phosphorylation of serine residues in histone H1 by the plant enzyme. Histone-related oligopeptides known to be substrates for animal histone kinases also served as substrates for the alfalfa kinase. Both of the studied peptides (GKKRKRSRKA; AAASFKAKK) inhibited phosphorylation of H1 histones by bovine and alfalfa kinases. The results of competition studies with the nonapeptide (AAASFKAKK), which is a PK-C specific substrate, suggest common features in target recognition between the plant Ca(2+)-dependent kinase and animal protein kinase C. We also propose that synthetic peptides like AAASFKAKK can be used as a tool to study substrates of plant kinases in crude cell extracts.

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Section: Discussionsupporting

confidence: 62%

Section: Introductionmentioning

confidence: 99%

The phosphorylation site of Ca2+-dependent protein kinase from alfalfa

et al. 1989

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“…The conformation of the substrate peptide/protein is also important for recognition by phosphorylase kinase (Tessmer & Graves, 1973; Tessmer et al, 1977;Viriya & Graves, 1979). A single basic residue (preferably arginine) on the carboxylor amino-terminal side of the target serine residue is the primary requirement for recognition by protein kinase C, although additional basic residues as well as structural features strongly influence the kinetics of peptide phosphorylation (Turner et al, 1985;Su et al, 1986;Kondo et al, 1987;House et al, 1987;Sakanoue et al, 1987). A similar requirement for basic residues is exhibited by myosin light chain kinase (Kemp et al, 1982).…”

Section: Discussionmentioning

confidence: 99%

Substrate recognition determinants for rhodopsin kinase: studies with synthetic peptides, polyanions, and polycations

Palczewski

Arendt²,

McDowell³

et al. 1989

Biochemistry

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Rhodopsin kinase phosphorylates serine- and threonine-containing peptides from bovine rhodopsin's carboxyl-terminal sequence. Km's for the peptides decrease as the length of the peptide is increased over the range 12-31 amino acids, reaching 1.7 mM for peptide 318-348 from the rhodopsin sequence. The Km for phosphorylation of rhodopsin is about 10(3) lower than that for the peptides, which suggests that binding of rhodopsin kinase to its substrate, photolyzed rhodopsin, involves more than just binding to the carboxyl-terminal peptide region that is to be phosphorylated. A synthetic peptide from the rhodopsin sequence that contains both serines and threonines is improved as a substrate by substitution of serines for the threonines, suggesting that serine residues are preferred as substrates. Analogous 25 amino acid peptides from the human red or green cone visual pigment, a beta-adrenergic receptor, or M1 muscarinic acetylcholine receptors are better substrates for bovine rhodopsin kinase than is the peptide from bovine rhodopsin. An acidic serine-containing peptide from a non-receptor protein, alpha s1B-casein, is also a good substrate for rhodopsin kinase. However, many basic peptides that are substrates for other protein kinases--histone IIA, histone IIS, clupeine, salmine, and a neurofilament peptide--are not phosphorylated by rhodopsin kinase. Polycations such as spermine or spermidine are nonessential activators of phosphorylation of rhodopsin or its synthetic peptide 324-348. Polyanions such as poly(aspartic acid), dextran sulfate, or poly(adenylic acid) inhibit the kinase. Poly(L-aspartic acid) is a competitive inhibitor with respect to rhodopsin (KI = 300 microM) and shows mixed type inhibition with respect to ATP.

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“…Although it is not clear whether this enzyme was protein kinase C or another protease-activated kinase reported by Traugh and her colleagues [35,361, phosphate incorporation into both serine residues was established by isoelectric focussing of radioactive tryptic peptides [34]. In the present study, phosphorylation of strate recognition by protein kinase C that at least one basic amino acid is arranged on either side of the phosphorylated site [21,37,39,41,441. The serine residue corresponding to Ser-9 in S6 protein was also proposed as the phosphorylated site of CAMP-dependent protein kinase [19].…”

Section: Discussionmentioning

confidence: 62%

“…Previous reports on the phosphorylated sites of various substrates for protein kinase C showed that basic amino acids existed around the phosphoserine or phosphothreonine residue [21,22,37,[39][40][41][42][43][44]. Studies on the substrate specificity of protein kinase C using synthetic peptides suggest that existence of a basic amino acid on the carboxyl-terminal side of the phosphorylated amino acid is important for decreasing the K , of the substrate and the specific recognition of phosphorylated site(s) for protein kinase C [21,37,43,441. This evidence coincided with the idea that Ser-5 is prefered to Ser-4 in peptide R' -A13 by protein kinase M and C [21].…”

Section: Phosphorylation Of Peptide R' -A13 By Protein Kinase M Andprmentioning

confidence: 99%

Comparative studies on phosphorylation of synthetic peptide analogue of ribosomal protein S6 and 40‐S ribosomal subunits between Ca^{2 +}/phospholipid‐dependent protein kinase and its protease‐activated form

Sakanoue

Hashimoto

Mizuta

et al. 1987

European Journal of Biochemistry

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showed essentially the same kinetic properties; V and apparent K , were determined to be 0.16 pmol min-' mg-' and 30 pM, respectively. At first, tryptic phosphopeptides were prepared at the early stage of phosphorylation and purified by high-performance liquid chromatography (HPLC). Through these analyses, four radioactive peptides were isolated. When protein kinase C was employed, phosphorylation was observed on all four peptides in a Ca2+/ phospholipid-dependent manner. Irrespective of the protein kinase employed, phosphate incorporation into these peptides increased linearly with time; the peptide concentration did not affect the ratio of phosphate distribution into these four peptides.Analysis of amino acid composition and phosphoamino acid of radioactive peptides obtained after extensive phosphorylation showed that phosphates were incorporated into Ser-4, Ser-5, Ser-9 and Ser-I 1. The latter three serine residues were major phosphorylated sites. When rat liver 40-S ribosomal subunits were employed as substrate for protein kinases C and M, a radioactive protein with I I~, , ,~~ = 31000, which corresponded to S6 protein, was detected on an autoradiogram of a sodium dodecyl sulfate/polyacrylamide slab gel. The rate of phosphorylation with protein kinase M was twice as fast as that with protein kinase C. The elution profile of radioactive tryptic peptides in HPLC suggest that phosphorylation occurred on the sites in S6 protein corresponding to Ser-5, Ser-9 and Ser-11 as major sites and Ser-4 as the minor one. These results indicate that protein kinase C has an ability to recognize at least four sites derived from hormone-dependent phosphorylation sites in ribosomal protein S6 irrespective of the mode of activation of this enzyme.Since the first description of Ca2 +/phospholipid-dependent protein kinase (protein kinase C) in rat brain by Takai et al. [I], experimental results have accumulated indicating that this enzyme is involved in various cellular functions and control of cell proliferation [2]. Reflecting the extensive research on this protein kinase, the number of functional proteins and enzymes proposed as substrate have been increasing over the past few years [2]. Phosphorylation of ribosomal protein S6 by protein kinase C was first reported by Le Peuch et al. [3] and described later by Parker et al. [4]. In parallel with these in vitro studies, phosphorylation of S6 protein was also observed in various cellular systems after treatment with tumor-promoting phorbol esters such as 12-0-tetradecanoylphorbol 13-acetate [5-91. It has been established that this phorbol ester activates protein kinase C by increasing the Abbreviations. R ' -A13, synthetic peptide Arg-Arg-Leu-Ser-SerLeu-Arg-Ala-Ser-Thr-Ser-Lys-Ala; P-Ser, phosphoserine; P-Thr, phosphothreonine.Enzymes. Protein kinase C (EC 2.7.

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Phosphorylation by protein kinase C of a synthetic heptapeptide bearing a lysine residue on the C terminal side of serine

Cited by 10 publications

References 22 publications

The phosphorylation site of Ca2+-dependent protein kinase from alfalfa

The phosphorylation site of Ca2+-dependent protein kinase from alfalfa

Substrate recognition determinants for rhodopsin kinase: studies with synthetic peptides, polyanions, and polycations

Comparative studies on phosphorylation of synthetic peptide analogue of ribosomal protein S6 and 40‐S ribosomal subunits between Ca^{2 +}/phospholipid‐dependent protein kinase and its protease‐activated form

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Phosphorylation by protein kinase C of a synthetic heptapeptide bearing a lysine residue on the C terminal side of serine

Cited by 10 publications

References 22 publications

The phosphorylation site of Ca2+-dependent protein kinase from alfalfa

The phosphorylation site of Ca2+-dependent protein kinase from alfalfa

Substrate recognition determinants for rhodopsin kinase: studies with synthetic peptides, polyanions, and polycations

Comparative studies on phosphorylation of synthetic peptide analogue of ribosomal protein S6 and 40‐S ribosomal subunits between Ca2 +/phospholipid‐dependent protein kinase and its protease‐activated form

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Comparative studies on phosphorylation of synthetic peptide analogue of ribosomal protein S6 and 40‐S ribosomal subunits between Ca^{2 +}/phospholipid‐dependent protein kinase and its protease‐activated form