scholarly journals Expression, purification and characterization of the ubiquitous protein kinase C-related kinase 1

1995 ◽  
Vol 309 (1) ◽  
pp. 315-320 ◽  
Author(s):  
R H Palmer ◽  
P J Parker
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Esters ◽  
Limited Proteolysis ◽  
Purification And Characterization ◽  
Kinase C ◽  
Apparent Homogeneity ◽  
Physiological Relevance ◽  
Sds Page

The recently described protein kinase C-related kinase (PRK) family is comprised of at least three members: PRK1, PRK2 and PRK3. Here the expression, purification and characterization of the ubiquitously expressed isoform, PRK1, is described. The enzyme was expressed in COS 7 cells and subsequently purified to apparent homogeneity by sequential column chromatography. The purified PRK1 protein migrates as a single 120 kDa polypeptide on SDS/PAGE. It displays a substrate specificity that in part resembles that of protein kinase C (PKC); however, unlike PKC, it is not activated by any combination of phorbol esters, diacylglycerol and Ca2+. Nevertheless, it can be activated by limited proteolysis, indicating a negative regulatory role for the N-terminal domain(s). PRK1 is also activated by phospholipids. The physiological relevance of this activation is discussed.

scholarly journals Purification and characterization of protein kinase C from the nematode Caenorhabditis elegans

1992 ◽  
Vol 282 (1) ◽  
pp. 219-223 ◽  
Author(s):  
T Sassa ◽  
J Miwa
Keyword(s):  
Protein Kinase C ◽  
Caenorhabditis Elegans ◽  
Protein Kinase ◽  
Column Chromatography ◽  
Purification And Characterization ◽  
C Elegans ◽  
Kinase C ◽  
Sds Page ◽  
Cytosolic Extract

Protein kinase C (PKC) of Caenorhabditis elegans was identified by enzymatic activity and [3H]phorbol 12,13-dibutyrate binding after DEAE-Sephacel column chromatography of a crude cytosolic extract. Ca(2+)-dependent activation of nematode PKC was observed in the presence of phosphatidylserine. The enzyme was maximally activated by 1,2-dioleoylglycerol or phorbol 12-myristate 13-acetate in the presence of phosphatidylserine and Ca2+. Hydroxyapatite column chromatography showed only one peak of PKC activity with histone H1 and myelin basic protein as substrates. The enzyme was purified to near homogeneity by sequential chromatography on polylysine-agarose and phosphatidylserine affinity columns. The purified protein showed a molecular mass of 79 kDa on SDS/PAGE. The substrate specificity of the C. elegans enzyme was shown to be different from that of mammalian PKCs. Here we describe some of the properties of the nematode enzyme.

Purification and Characterization of Protein Kinase C from a Higher Plant, Brassica campestris L

1994 ◽  
Vol 203 (1) ◽  
pp. 311-318 ◽  
Author(s):  
T. Nanmori ◽  
W. Taguchi ◽  
M. Kinugasa ◽  
Y. Oji ◽  
S. Sahara ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Brassica Campestris ◽  
Purification And Characterization ◽  
Higher Plant ◽  
Kinase C

Isolation and characterization of PKC-L, a new member of the protein kinase C-related gene family specifically expressed in lung, skin, and heart

1991 ◽  
Vol 11 (1) ◽  
pp. 126-133 ◽  
Author(s):  
N Bacher ◽  
Y Zisman ◽  
E Berent ◽  
E Livneh
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Gene Family ◽  
Phorbol Esters ◽  
Structural Features ◽  
Cos Cells ◽  
Isolation And Characterization ◽  
Kinase C ◽  
Human Epidermoid Carcinoma

We have isolated and characterized a new human cDNA, coding for a protein kinase, related to the protein kinase C (PKC) gene family. Although this protein kinase shares some homologous sequences and structural features with the four members of the PKC family initially isolated (alpha, beta I, beta II, and gamma), it shows more homology with the recently described PKC-related subfamily, encoded by the cDNAs delta, epsilon, and zeta. The transcript for this gene product, termed PKC-L, is most abundant in lung tissue, less expressed in heart and skin tissue, and exhibited very low expression in brain tissue. Thus, its tissue distribution is different from that described for other mammalian members of the PKC gene family, their expression being enriched in brain tissues. PKC-L is also expressed in several human cell lines, including the human epidermoid carcinoma line A431. The ability of phorbol esters to bind to and stimulate the kinase activity of PKC-L was revealed by introducing the cDNA into COS cells.

scholarly journals A growth factor-repressible gene associated with protein kinase C-mediated inhibition of adipocyte differentiation.

1988 ◽  
Vol 107 (1) ◽  
pp. 279-286 ◽  
Author(s):  
M Navre ◽  
G M Ringold
Keyword(s):  
Protein Kinase C ◽  
Growth Factor ◽  
Protein Kinase ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Phorbol Esters ◽  
Adipocyte Differentiation ◽  
Fibroblast Growth ◽  
Kinase C

The conversion of determined adipoblasts to fully differentiated adipocytes requires appropriate environmental conditions. A strict dependence on cell confluence and a facilitation by glucocorticoid hormones have previously been described. We have found that agents that are capable of activating protein kinase C, such as basic fibroblast growth factor and phorbol esters, inhibit the differentiation of the adipogenic cell line TA1 without stimulating cell growth. Here we describe the sequence and characterization of a cDNA (clone 5) that detects an RNA, the expression of which is enhanced by glucocorticoids and increasing cell density. In contrast, activators of protein kinase C including basic fibroblast growth factor, phorbol esters, and synthetic diacylglycerols inhibit clone 5 gene expression. It appears that clone 5 expression is closely linked to environmental and hormonal factors that promote the differentiation of adipogenic cells.

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