Abstract:
:Ribose-modified fluorescent nucleotide analogs, 3'-O-anthraniloyl and 3'-O-(N-methylanthraniloyl) derivatives of AT(D)P, dAT(D)P, CT(D)P, UT(D)P, IT(D)P, and GT(D)P, were synthesized for use as substrates and affinity labels for the myosin ATPase [Hiratsuka, T. (1983) Biochim. Biophys. Acta 742, 496-508]. None of the fluorescent nucleoside triphosphate (NTP) analogs was significantly different from the corresponding natural NTP in its ability to support superprecipitation of actomyosin. When fluorescent and natural NTPs were used as substrates for the myosin subfragment-1(S-1) ATPase in the presence of 1mM vanadate ion (V1), a slight initial inhibition of the S-1 NTPase was followed by progressive inhibition to more than 60% over a period of 1 h. The apparent second-order rate constants were 0.14-0.44M-1 . s-1, suggesting the formation of the inactive fluorescent NDP-labeled S-1. After incubation of S-1 with the nucleoside diphosphate (NDP) analog in the presence of Vi, the resultant fluorescent NDP-labeled S-1 was isolated free of unbound Vi and the analog by gel filtration. The isolated complexes had stoichiometries of 0.6-1.1 NDP analog per S-1 active site. Native polyacrylamide gel electrophoresis revealed conveniently that the NDP analog is associated with S-1 as indicated by two intense fluorescent bands corresponding to S-1 isozymes. On dissociating gels, the analog was released from S-1, suggesting that the labeled S-1 is held together by strong secondary forces rather than covalent bonds.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Hiratsuka Tdoi
10.1093/oxfordjournals.jbchem.a134806subject
Has Abstractpub_date
1984-07-01 00:00:00pages
147-54issue
1eissn
0021-924Xissn
1756-2651journal_volume
96pub_type
杂志文章abstract::Sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) are blood-borne lysophospholipids with a wide spectrum of biological activities, which include stimulation of cell growth, prevention of apoptosis, regulation of actin cytoskeleton, and modulation of cell shape, cell migration, and invasion. Activated plate...
journal_title:Journal of biochemistry
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journal_title:Journal of biochemistry
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abstract::A DNA polymerase which was active with unprimed poly(dC) as a template was detected in the DNA polymerase alpha fraction from rat thymoma tissue. The activity was dependent on rGTP and associated with ribonucleotide polymerizing activity. The activity was partially sensitive to aphidicolin and dideoxy GTP and resistan...
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更新日期:1983-01-01 00:00:00
abstract::Activation-induced cell death (AICD) plays a critical role in the maintenance of homeostasis and peripheral tolerance in the immune system, and is mediated by Fas ligand (FasL) expression and the interaction between Fas and FasL. In the present study, we examined the role of the ubiquitin-proteasome system in AICD usi...
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doi:10.1093/oxfordjournals.jbchem.a133081
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pub_type: 杂志文章,评审
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更新日期:1994-04-01 00:00:00
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更新日期:1994-02-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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journal_title:Journal of biochemistry
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更新日期:2021-01-21 00:00:00
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journal_title:Journal of biochemistry
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doi:10.1093/oxfordjournals.jbchem.a132539
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a135481
更新日期:1986-01-01 00:00:00
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更新日期:1994-03-01 00:00:00
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journal_title:Journal of biochemistry
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更新日期:1991-12-01 00:00:00