کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
9573486 1388902 2005 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
MgATP-induced conformational change of the catalytic subunit of cAMP-dependent protein kinase
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی تئوریک و عملی
پیش نمایش صفحه اول مقاله
MgATP-induced conformational change of the catalytic subunit of cAMP-dependent protein kinase
چکیده انگلیسی
Conformational changes of the cAMP-dependent protein kinase (PKA) catalytic (C) subunit are critical for the catalysis of γ-phosphate transfer from adenosine 5′-triphosphate (ATP) to target proteins. Time-resolved fluorescence anisotropy (TRFA) was used to investigate the respective roles of Mg2+, ATP, MgATP, and the inhibitor peptide (IP20) in the conformational changes of a 5,6-carboxyfluorescein succinimidyl ester (CF) labeled C subunit (CFC). TRFA decays were fit to a biexponential equation incorporating the fast and slow rotational correlation times ϕF and ϕS. The CFC apoenzyme exhibited the rotational correlation times ϕF=1.8±0.3 ns and ϕS=20.1±0.6 ns which were reduced to ϕF=1.1±0.2 ns and ϕS=13.3±0.9 ns in the presence of MgATP. The reduction in rotational correlation times indicated that the CFC subunit adopted a more compact shape upon formation of a CFC·MgATP binary complex. Neither Mg2+ (1-3 mM) nor ATP (0.4 mM) alone induced changes in the CFC subunit conformation equivalent to those induced by MgATP. The effect of MgATP was removed in the presence of ethylenediaminetetraacetic acid (EDTA). The addition of IP20 and MgATP to form the CFC·MgATP·IP20 ternary complex produced rotational correlation times similar to those of the CFC·MgATP binary complex. However, IP20 alone did not elicit an equivalent reduction in rotational correlation times. The results indicate that binding of MgATP to the C subunit may induce conformation changes in the C subunit necessary for the proper stereochemical alignment of substrates in the subsequent phosphorylation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biophysical Chemistry - Volume 113, Issue 2, 1 February 2005, Pages 193-199
نویسندگان
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