Heterogeneity of protein kinase NII from rat liver nuclei

Inoue, Akira; Tei, Yoko; Qi, Shangle; Higashi, Youtaro; Yukioka, Munehiko; Morisawa, Seiji

doi:10.1016/0006-291x(84)90427-3

Cited by 25 publications

(7 citation statements)

References 16 publications

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“…They displayed several nonclassical chromatographic, physical, and biochemical characteristics, summarized in Table 4. Isozyme forms (Litchfield et al, 1990) and heterogeneity (Inoue et al, 1984) of CK II have been reported. For example, two forms of protein kinase NII (the nuclear form of CK II), possessing the same molecular weight, subunit composition, and identical K, for ATP, but different elution profiles and reaction kinetics for casein, were purified from rat liver nuclei (Inoue et al, 1984) .…”

Section: Discussionmentioning

confidence: 99%

“…Isozyme forms (Litchfield et al, 1990) and heterogeneity (Inoue et al, 1984) of CK II have been reported. For example, two forms of protein kinase NII (the nuclear form of CK II), possessing the same molecular weight, subunit composition, and identical K, for ATP, but different elution profiles and reaction kinetics for casein, were purified from rat liver nuclei (Inoue et al, 1984) . Although it was well established that a and a' subunits derived from separate genes, it remained unclear whether they perform distinct functions (Padmanabha and Glover, 1987;Litchfield et al, 1990;Maridor et al, 1991) .…”

Section: Discussionmentioning

confidence: 99%

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Purification and Characterization of Two Casein Kinase Type II Isozymes from Bovine Brain Gray Matter

Mitev¹,

Pauloin²,

Houdebiné³

1994

Journal of Neurochemistry

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Highly purified casein kinase II (CK II) isozymes from bovine brain gray matter (BBGM) were obtained by means of a new purification procedure consisting of one phosphocellulose and three Mono‐Q steps. The phosphocellulose eluate showed two BBGM‐CK II activities. The first minor component (BBGM‐CK IIa) was eluted with 0.9 M NaCl and the major component was eluted at 1.1 M NaCl (BBGM‐CK IIb). The protein complexes responsible for these two activities were comprised of three subunits, i.e., α (40 kDa), α′ (38 kDa), and β (28 kDa), with various subunit ratios. The two isozymes displayed the same behavior on Superose 12 fast protein liquid chromatographic gel filtration and sucrose density centrifugation. BBGM‐CK IIa and b showed chromatographic and biochemical differences including differing Km for ATP and GTP and Ki for heparin and 2,3‐bisphosphoglycerate. The properties of the main peak (BBGM‐CK IIb) were studied in detail. The stimulatory effect of Mg2+, Mn2+, and Co2+ was highly dependent both on the nature of the substrate and on ionic type and concentration. It is surprising that with phosvitin as substrate, BBGM‐CK IIb was fully active even in the absence of Mg2+ and NaCl. The inhibitory effect of heparin and the stimulatory effects of NaCl, KCl, spermine, and polylysine were highly dependent on the ionic strength, buffer type, and substrate. BBGM‐CK II isozymes phosphorylated stathmine in the presence of polylysine, but the requirement for polybasic compounds was not absolute, as is the case with calmodulin and clathrin β‐light chain. The unusual chromatographic behavior and biochemical properties of these BBGM‐CK II isozymes, compared with the classical CK II, could be explained at least in part by their subunit ratios.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of Two Casein Kinase Type II Isozymes from Bovine Brain Gray Matter

Mitev¹,

Pauloin²,

Houdebiné³

1994

Journal of Neurochemistry

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“…Unoccupied receptors were prepared with Lipidex as described above. The occupied and unoccupied receptors were eluted with a 50-400 mM NaCl linear gradient in buffer T on parallel columns of DEAE-Sephadex A-50 as described elsewhere [10].…”

Section: Deae-sephadex A-50 Column Chromatographymentioning

confidence: 99%

“…Unoccupied receptors (A) prepared by incubation of a nuclear extract with 50 uM oleoyl-CoA and passage through a Lipidex column were eluted from DEAE-Sephadex A-50 (8 x 40 mm) with a 50-400 mM NaCI gradient in buffer T. Occupied receptors (0) prepared by incubation of the nuclear extract with ['251]T3 were eluted at the same time on a parallel column (10). Fractions of 0.8 ml were collected.…”

Section: T3-binding Parameters Of the Unoccupied Receptormentioning

confidence: 99%

Preparation of unoccupied thyroid-hormone receptor

Inoue

1994

Biochemical Journal

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Thyroid hormone (3,5,3'-tri-iodothyronine; T3) regulates gene expression through binding to its specific receptor in the nucleus. In euthyroid animals, roughly half of all receptors are occupied by the hormone. Nuclear extracts thus yield mixtures of occupied and unoccupied receptors. We present here a simple method for transforming occupied receptors into unoccupied ones. In vitro, the T3-receptor complex dissociated in a half-dissociation time exceeding 100 h at 0 degrees C, and at temperatures that accelerated the dissociation the receptor was quickly inactivated. Long-chain-fatty-acyl-CoAs, on the other hand, greatly accelerated the dissociation of T3-receptor complex at 0 degree C. The receptor was extracted from rat liver nuclei, incubated with oleoyl-CoA to release the bound hormone, and passed through a small column of Lipidex, which strongly adsorbed both oleoyl-CoA and the dissociated hormone. The receptor was recovered in the flow-through fraction in its unoccupied form, as seen by the results of DEAE-Sephadex column chromatography and the loss of all previously bound [125I]T3. The maximum T3-binding capacity of the unoccupied receptor was about 1.5-fold that of the untreated sample, and the dissociation constant was unaltered. The results suggest that most nuclear thyroid-hormone receptors occupied by the hormone were transformed into unoccupied ones. From the T3-binding capacity before and after oleoyl-CoA treatment, the in vivo T3 occupancy of the receptor was estimated. The procedure is easy to perform, and the method should be useful for studies of unoccupied receptors.

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“…In rat liver, NI appears to consist of two subunits of molecular weight 25 kDa (161), and Nil is tetrameric (aW/3J with a molecular weight of about 130-135 kDa (162), although even Nil shows heterogeneity (163).…”

Section: Potential Role Of Protein Phosphorylation In Rna Transportmentioning

confidence: 99%

Nucleocytoplasmic RNA transport

1985

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A number of closely related post-transcriptional facets of RNA metabolism show nuclear compartmentation, including capping, methylation, splicing reactions, and packaging in ribonucleoprotein particles (RNP). These nuclear 'processing' events are followed by the translocation of the finished product across the nuclear envelope. Due to the inherent complexity of these interrelated events, in vitro systems have been designed to examine the processes separately, particularly so with regard to translocation. A few studies have utilized nuclear transplantation/microinjection techniques and specialized systems to show that RNA transport occurs as a regulated phenomenon. While isolated nuclei swell in aqueous media and dramatic loss of nuclear protein is associated with this swelling, loss of RNA is not substantial, and most studies on RNA translocation have employed isolated nuclei. The quantity of RNA transported from isolated nuclei is related to hydrolysis of high-energy phosphate bonds in nucleotide additives. The RNA is released predominantly in RNP: messenger-like RNA is released in RNP which have buoyant density and polypeptide composition similar to cytoplasmic messenger RNP, but which have distinctly different composition from those in heterogeneous nuclear RNP. Mature 18 and 28S ribosomal RNA is released in 40 and 60S RNP which represent mature ribosomal subunits. RNA transport proceeds with characteristics of an energy-requiring process, and proceeds independently of the presence or state of fluidity of nuclear membranes. The energy for transport appears to be utilized by a nucleoside triphosphatase (NTPase) which is distributed mainly within heterochromatin at the peripheral lamina. Photoaffinity labeling has identified the pertinent NTPase as a 46 kD polypeptide which is associated with nuclear envelope and matrix preparations. The NTPase does not appear to be modulated via direct phosphorylation or to reflect kinase-phosphatase activities. A large number of additives (including RNA and insulin) produce parallel effects upon RNA transport and nuclear envelope NTPase, strengthening the correlative relationship between these activities. Of particular interest has been the finding that carcinogens induce specific, long-lasting increases in nuclear envelope (and matrix) NTPase; this derangement may underlie the alterations in RNA transport associated with cancer and carcinogenesis.(ABSTRACT TRUNCATED AT 400 WORDS)

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Heterogeneity of protein kinase NII from rat liver nuclei

Cited by 25 publications

References 16 publications

Purification and Characterization of Two Casein Kinase Type II Isozymes from Bovine Brain Gray Matter

Purification and Characterization of Two Casein Kinase Type II Isozymes from Bovine Brain Gray Matter

Preparation of unoccupied thyroid-hormone receptor

Nucleocytoplasmic RNA transport

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