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41.
42.
The polyamine-derived amino acid hypusine: its post-translational formation in eIF-5A and its role in cell proliferation 总被引:1,自引:0,他引:1
Myung Hee Park Young Ae Joe Kee Ryeon Kang Young Bok Lee Edith C. Wolff 《Amino acids》1996,10(2):109-121
Summary The unusual amino acid hypusine [N
-(4-amino-2-hydroxybutyl)lysine] is a unique component of one cellular protein, eukaryotic translation initiation factor 5A (eIF-5A, old terminology, eIF-4D). It is formed posttranslationally and exclusively in this protein in two consecutive enzymatic reactions, (i) modification of a single lysine residue of the eIF-5A precursor protein by the transfer of the 4-aminobutyl moiety of the polyamine spermidine to its-amino group to form the intermediate, deoxyhypusine [N
-(4-aminobutyl)lysine] and (ii) subsequent hydroxylation of this intermediate to form hypusine. The amino acid sequences surrounding the hypusine residue are strictly conserved in all eukaryotic species examined, suggesting the fundamental importance of this amino acid throughout evolution. Hypusine is required for the activity of eIF-5Ain vitro. There is strong evidence that hypusine and eIF-5A are vital for eukaryotic cell proliferation. Inactivation of both of the eIF-5A genes is lethal in yeast and the hypusine modification appears to be a requirement for yeast survival (Schnier et al., 1991 [Mol Cell Biol 11: 3105–3114]; Wöhl et al., 1993 [Mol Gen Genet 241: 305–311]). Furthermore, inhibitors of either of the hypusine biosynthetic enzymes, deoxyhypusine synthase or deoxyhypusine hydroxylase, exert strong anti-proliferative effects in mammalian cells, including many human cancer cell lines. These inhibitors hold potential as a new class of anticancer agents, targeting one specific eukaryotic cellular reaction, hypusine biosynthesis. 相似文献
43.
Microbial flocculant from Arcuadendron sp. TS-49 总被引:9,自引:0,他引:9
Summary A flocculant was purified from the culture broth of Archuadendron sp. TS-49 by a series of precipitations with acetone, 60% ammonium sulfate-butanol, salting-out by dialysis, and cetylpyridinium chloride. The flocculating activity was observed most highly at pH 3.0 and markedly enhanced by the addition of salts, especially in the case of FeCl3 or FeSO4. This bioflocculant efficiently flocculated all tested solids, including various microorganisms and organic/ inorganic materials. Qualitative analyses of the bioflocculant showed that it might contain hexosamine, uronic acid, neutral sugar, and protein. 相似文献
44.
Weidong Jiang Moon-Young Lim Hye-Joo Yoon Jeremy Thorner G. Steven Martin John Carbon 《Molecular & general genetics : MGG》1995,246(3):360-366
We find that overexpression in yeast of the yeast MCK1 gene, which encodes a meiosis and centromere regulatory kinase, suppresses the temperature-sensitive phenotype of certain mutations in essential centromere binding protein genes CBF2 and CBF5. Since Mck1p is a known serine/threonine protein kinase, this suppression is postulated to be due to Mck1p-catalyzed in vivo phosphorylation of centromere binding proteins. Evidence in support of this model was provided by the finding that purified Mck1p phosphorylates in vitro the 110 kDa subunit (Cbf2p) of the multimeric centromere binding factor CBF3. This phosphorylation occurs on both serine and threonine residues in Cbf2p. 相似文献
45.
We used quantitative complementation assays to characterize individual DNA polymerase (Pol) mutants for their ability to function in DNA replication and DNA repair. We also describe a screen for detecting imitator activity of DNA polymerase mutants. By using these bioassays, together with DNA polymerase activity gels, we characterized 15 new DNA polymerase mutants that display a wide spectrum of phenotypes. Most of these mutants are generally defective in their ability to synthesize DNA. However, two of our Pol mutants show more complex phenotypes: they are able to function in DNA repair but unable to participate in DNA replication. One of our mutants displays imitator activity in vivo. Our work provides a model to study mutant mammalian enzymes inEscherichia coli with phenotypes that are otherwise difficult to assess. 相似文献
46.
The Old protein of bacteriophage P2 is responsible for interference with the growth of phage lambda and for killing of recBC mutant Escherichia coli. We have purified Old fused to the maltose-binding protein to 95% purity and characterized its enzymatic properties. The Old protein fused to maltose-binding protein has exonuclease activity on double-stranded DNA as well as nuclease activity on single-stranded DNA and RNA. The direction of digestion of double-stranded DNA is from 5' to 3', and digestion initiates at either the 5'-phosphoryl or 5'-hydroxyl terminus. The nuclease is active on nicked circular DNA, degrades DNA in a processive manner, and releases 5'-phosphoryl mononucleotides. 相似文献
47.
Hyung-Joo Jin Jeong–Ha Kim Chul Hyun Sohn R.E. DeWreede Tae–Joo Choi G.H.N. Towers J.B. Hudson Yong–Ki Hong 《Journal of applied phycology》1997,9(4):383-388
Fifty-nine species of marine macrophytes from the coasts of British Columbia, Canada and Korea have been screened for the
presence of PCR inhibitors, namely inhibitors of Taq DNA polymerase. Eleven of the species displayed some inhibitor activity.
At the concentration of 5 μg of methanol extract in 25μL reaction mixture of PCR containing 1.5 unit of Taq DNA polymerase,
one (Ulva sp.) of 8 Chlorophyta, eight (Colpomenia bullosa, Ecklonia cava, Endarachne binghamiae, Fucus distichus, Hizikia
fusiformis, Sargassum confusum, Sargassum sagamianum, and Sargassum thunbergii) of 28 Phaeophyta, and one (Symphyocladia latiuscula)
of 34 Rhodophyta showed inhibition in PCR amplification. In the case of the water extract, two (Cladophora columbiana, Ulva
sp.) Chlorophyta, seven (Endarachne binghamiae, Fucus distichus, Hizikia fusiformis, Sargassum confusum, Sargassum sagamianum,
Sargassum horneri, Scytosiphon dotyi) Phaeophyta, no Rhodophyta and one (Phyllospadix scouleri) seagrass showed inhibition
in PCR amplification. the methanol fraction of Sargassum confusum and the water fraction of Fucus gardneri (mid–intertidal)
have been found to inhibit PCR at level as low as 0.5 μg in 25μL of PCR reaction mixture.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
48.
Genetic variation of herpesvirus saimiri subgroup A transforming protein and its association with cellular src. 总被引:2,自引:2,他引:0 下载免费PDF全文
H Lee J J Trimble D W Yoon D Regier R C Desrosiers J U Jung 《Journal of virology》1997,71(5):3817-3825
Herpesvirus saimiri strain 11 of subgroup A contains a gene called the saimiri transformation-associated protein, STP, which is not required for viral replication but is required for in vitro immortalization and for the lymphoma-inducing capacity of the virus. To assess the effects of sequence variation on STP function, STP genes from six subgroup A isolates were cloned and sequenced. Sequence comparisons revealed extensive amino acid substitutions within the central region, but the acidic amino terminus and the hydrophobic carboxyl terminus were well conserved. Amino acid identities varied from 73 to 99% among all two-way comparisons. The highly conserved YAEV/I motif at amino acid residues 115 to 118 was preceded by negatively charged glutamic acid residues and thus matched very well the consensus sequence for binding to SH2 domains of src family kinases. The STPs of these subgroup A strains were shown to associate with cellular src and to be an in vitro substrate for src kinase. Mutational analysis of STP-A11 showed that binding to src kinase required the tyrosine residue at 115, showing that YAEV/I is a likely binding motif for src. Also, tyrosine phosphorylation of STP-A11 by src led to subsequent binding to lck and fyn in vitro. Thus, the association of STP with src is likely to be important for T-cell transformation by subgroup A strains of herpesvirus saimiri. 相似文献
49.
Possible role of macrophage-derived soluble mediators in the pathogenesis of encephalomyocarditis virus-induced diabetes in mice. 总被引:4,自引:1,他引:3 下载免费PDF全文
K Hirasawa H S Jun K Maeda Y Kawaguchi S Itagaki T Mikami H S Baek K Doi J W Yoon 《Journal of virology》1997,71(5):4024-4031
Pancreatic islets from DBA/2 mice infected with the D variant of encephalomyocarditis (EMC-D) virus revealed lymphocytic infiltration with moderate to severe destruction of pancreatic beta cells. Our previous studies showed that the major population of infiltrating cells at the early stages of infection is macrophages. The inactivation of macrophages prior to viral infection resulted in the prevention of diabetes, whereas activation of macrophages prior to viral infection resulted in the enhancement of beta-cell destruction. This investigation was initiated to determine whether macrophage-produced soluble mediators play a role in the destruction of pancreatic beta cells in mice infected with a low dose of EMC-D virus. When we examined the expression of the soluble mediators interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and inducible nitric oxide synthase (iNOS) in the pancreatic islets, we found that these mediators were clearly expressed at an early stage of insulitis and that this expression was evident until the development of diabetes. We confirmed the expression of these mediators by in situ hybridization with digoxigenin-labelled RNA probes or immunohistochemistry in the pancreatic islets. Mice treated with antibody against IL-1beta or TNF-alpha or with the iNOS inhibitor aminoguanidine exhibited a significant decrease in the incidence of diabetes. Mice treated with a combination of anti-IL-1beta antibody, anti-TNF-alpha antibody, and aminoguanidine exhibited a greater decrease in the incidence of disease than did mice treated with one of the antibodies or aminoguanidine. On the basis of these observations, we conclude that macrophage-produced soluble mediators play an important role in the destruction of pancreatic beta cells, resulting in the development of diabetes in mice infected with a low dose of EMC-D virus. 相似文献
50.
Phosphorylation of rat muscle acetyl-CoA carboxylase by AMP-activated protein kinase and protein kinase A 总被引:7,自引:0,他引:7
Winder W. W.; Wilson H. A.; Hardie D. G.; Rasmussen B. B.; Hutber C. A.; Call G. B.; Clayton R. D.; Conley L. M.; Yoon S.; Zhou B. 《Journal of applied physiology》1997,82(1):219-225
Winder, W. W., H. A. Wilson, D. G. Hardie, B. B. Rasmussen,C. A. Hutber, G. B. Call, R. D. Clayton, L. M. Conley, S. Yoon, and B. Zhou. Phosphorylation of rat muscle acetyl-CoA carboxylase byAMP-activated protein kinase and protein kinase A. J. Appl. Physiol. 82(1): 219-225, 1997This studywas designed to compare functional effects of phosphorylation of muscleacetyl-CoA carboxylase (ACC) by adenosine 3,5-cyclicmonophosphate-dependent protein kinase (PKA) and by AMP-activatedprotein kinase (AMPK). Muscle ACC (272 kDa) was phosphorylated and thensubjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresisfollowed by autoradiography. Functional effects of phosphorylation weredetermined by measuring ACC activity at different concentrations ofeach of the substrates and of citrate, an activator of the enzyme. Themaximal velocity(Vmax) and theMichaelis constants(Km) for ATP,acetyl-CoA, and bicarbonate were unaffected by phosphorylation by PKA.Phosphorylation by AMPK increased theKm for ATP andacetyl-CoA. Sequential phosphorylation by PKA and AMPK, first withoutlabel and second with label, appeared to reduce the extent of label incorporation, regardless of the order. The activation constant (Ka) forcitrate activation was increased to the same extent by AMPKphosphorylation, regardless of previous or subsequent phosphorylation by PKA. Thus muscle ACC can be phosphorylated by PKA but with noapparent functional effects on the enzyme. AMPK appears to be the moreimportant regulator of muscle ACC. 相似文献