the structural basis of the cross-bridge cycle
DESCRIPTION
The structural basis of the cross-bridge cycle. DECORATED ACTIN was the first em reconstruction Moore, Huxley, and DeRosier 1970 J. Mol Biol. Reconstruction by filtered least squares. Projection every 1° (181 projections) Number of observational equations is 161 x 181=29141 - PowerPoint PPT PresentationTRANSCRIPT
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The structural basis of the cross-bridge cycle
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DECORATED ACTIN
was the first em reconstruction
Moore, Huxley, and DeRosier
1970 J. Mol Biol.
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Reconstruction by filtered least squares
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P f = g
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The classical solution to the equations
P f= g + e
which minimises the sum of squares oferror seTe is
PT P f = PTg
PT P is th enorma l o r Hessia n matrix
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The solution is
f = (PTP)-1 PTg
but PTg is the back projection.
Thus the inverse least squares matrix (PTP)-1
provides the analytical filter for turning theback projection into the required density
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Projection every 1° (181 projections)
Number of observational equations is
161 x 181=29141
Order of normal matrix 17645.
Problem should be well determined
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Oops!!
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Crowther, R. A., DeRosier, D. J. and Klug, A. (1972) The reconstruction of a three-dimensional structure from projections and its application to electron microscopy. Proc. Royal Soc. London 317A: 319-40.
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Case 2. Projection every 3°, not enough data to invert the normal matrix. How much data is there?
To determine the rank of a positive (semi) definite matrix we determine its eigenvalue spectrum
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Decorated actin at 13Å resolving power calculated by filtered least squares
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Electron cryo-microscopy (of decorated actin) shows how strong binding of myosin to actin releases nucleotide Holmes, Angert, Kull, Jahn, Schroeder (2003) Nature 425, 423-427
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Four sub-domain refinement
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The cross-bridge cycle
As revealed by crystallography
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Geeves, M.A. & Holmes, K.C. (2005)
The molecular mechanism of muscle contraction
Adv Protein Chem 71, 161-93
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Lymn & Taylor, 1971
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The myosin motor domain must move the lever arm from pre- to
post- power stroke.
It must bind to actin strongly or weakly
Both functions are controlled by ATP binding and hydrolysis
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Crystallography on the motordomain (particularly Dictyostelium Myosin 2) shows two conformations of myosin:
lever UP and lever DOWN
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Diskussionsthemen• Nennen Sie die Hauptthesen,
über die Sie sprechen werden.
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Thema 1• Details über das Thema
• Argumente und Beispiele
• Warum ist das Thema für Ihr Publikum von Bedeutung?
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switch 1
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QuickTime™ and aApple Motion JPEG Format A decompressor
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Fischer et al (2005) PNAS102 6873-8
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Fischer et al (2005) PNAS102 6873-8
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Weak and Strong Binding
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Coureux, P. D.Wells, A. L.Menetrey, J.Yengo, C. M.Morris, C. A.Sweeney, H. L.Houdusse, A. (2003)
A structural state of the myosin V motor without bound nucleotide
Nature 425 419-23
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Rayment_chicken
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Upper/lower 50K fitted
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Myo 5
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?
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