Persistence Length of Human Cardiac α-Tropomyosin Measured by Single Molecule Direct Probe MicroscopyReportar como inadecuado

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α-Tropomyosin αTm is the predominant tropomyosin isoform in adult human heart and constitutes a major component in Ca2+-regulated systolic contraction of cardiac muscle. We present here the first direct probe images of WT human cardiac αTm by atomic force microscopy, and quantify its mechanical flexibility with three independent analysis methods. Single molecules of bacterially-expressed human cardiac αTm were imaged on poly-lysine coated mica and their contours were analyzed. Analysis of tangent-angle θs correlation along molecular contours, second moment of tangent angles <θ2s>, and end-to-end length Le-e distributions respectively yielded values of persistence length Lp of 41–46 nm, 40–45 nm, and 42–52 nm, corresponding to 1–1.3 molecular contour lengths Lc. We also demonstrate that a sufficiently large population, with at least 100 molecules, is required for a reliable Lp measurement of αTm in single molecule studies. Our estimate that Lp for αTm is only slightly longer than Lc is consistent with a previous study showing there is little spread of cooperative activation into near-neighbor regulatory units of cardiac thin filaments. The Lp determined here for human cardiac αTm perhaps represents an evolutionarily tuned optimum between Ca2+ sensitivity and cooperativity in cardiac thin filaments and likely constitutes an essential parameter for normal function in the human heart.

Autor: Campion K. P. Loong , Huan-Xiang Zhou, P. Bryant Chase



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