کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
607942 880564 2013 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A visualized observation of calcium-dependent gelsolin activity upon the surface coverage of fluorescent-tagged actin filaments
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی شیمی کلوئیدی و سطحی
پیش نمایش صفحه اول مقاله
A visualized observation of calcium-dependent gelsolin activity upon the surface coverage of fluorescent-tagged actin filaments
چکیده انگلیسی

Gelsolin regulates the dynamics of F-actin by binding to F-actin to sever and cap. In the present study, a novel approach is introduced to observe gelsolin activity through the coverage of surface-bound F-actin. Gelsolin was immobilized on streptavidin coated surface using biotinylation and, as a result, the interaction between gelsolin and F-actin was visualized. Consequently, the coverage of F-actin reflects the activity of gelsolin as a function of free Ca2+ concentrations. In order to prevent non-specific binding of F-actin, the combinations of BSA and Tween-20 as blocking agents were investigated. Moreover, the measurement of the length of F-actin with actin–gelsolin mixtures at various ratios provided the verification of gelsolin activity after biotinylation. The data shows the increase in Ca2+ concentration leads to a proportional increase in F-actin coverage, giving to half-maximal coverage at ~2.9 μM. Furthermore, the length of bound F-actin was found to decrease along with increasing Ca2+ concentration, and full-length F-actin was rarely observed. This may suggest that severing and capping activities of gelsolin occur without more additional Ca2+ for subsequent activation after full-length gelsolin binds to a side of F-actin. This finding may provide a key to understand gelsolin activity.

Figure optionsDownload high-quality image (97 K)Download as PowerPoint slideHighlights
► A heterogeneous assay is developed to study gelsolin activity using F-actin.
► Blocking of non-specific binding sites is performed for the reliability of assay.
► The half-maximal coverage of F-actin occurred at 2.9 μM of free Ca2+.
► Severing and capping occurs without addition of Ca2+ after binding to F-actin.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Colloid and Interface Science - Volume 389, Issue 1, 1 January 2013, Pages 182–187
نویسندگان
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