Visualizing filamentous actin on lipid bilayers by atomic force microscopy in solution

The surface structure of actin filaments (F‐actin) was visualized at high resolution, by atomic force microscopy (AFM) in aqueous solution, in large paracrystals prepared on positively charged lipid monolayers. The increased stability of these closely packed specimens allowed us to show that both the long pitch (38 nm) and the monomer (5.8 nm) can be directly resolved by AFM in the contact mode. The right‐handed helical surface, distinguishable in high resolution images, was compared with reconstructed models based on electron microscopy. The height of the rafts, a measure of the actin filament diameter, was 10 ± 1 nm, whereas the smaller inter‐filament distance, 8 ± 1 nm, was consistent with interdigitation of the filaments. The 10 ± 1 nm F‐actin diameter is in good agreement with the results of fibre X‐ray diffraction. As such specimens are relatively easy to prepare without specialized equipment, this method may allow the study of the thin filaments in which F‐actin‐associated proteins are also present.

[1]  D. DeRosier,et al.  How to analyze electron micrographs of rafts of actin filaments crosslinked by actin-binding proteins. , 1998, Journal of molecular biology.

[2]  G Büldt,et al.  Surface structures of native bacteriorhodopsin depend on the molecular packing arrangement in the membrane. , 1999, Journal of molecular biology.

[3]  Z. Shao,et al.  New approach for atomic force microscopy of membrane proteins. The imaging of cholera toxin. , 1993, Journal of molecular biology.

[4]  D. St-Onge,et al.  Evidence of direct interaction between actin and membrane lipids. , 1989, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[5]  M. Borgnia,et al.  High resolution AFM topographs of the Escherichia coli water channel aquaporin Z , 1999, The EMBO journal.

[6]  D. Czajkowsky,et al.  The vacuolating toxin from Helicobacter pylori forms hexameric pores in lipid bilayers at low pH. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[7]  B Honig,et al.  Electrostatic binding of proteins to membranes. Theoretical predictions and experimental results with charybdotoxin and phospholipid vesicles. , 1997, Biophysical journal.

[8]  A. Engel,et al.  High resolution imaging of native biological sample surfaces using scanning probe microscopy. , 1997, Current opinion in structural biology.

[9]  Gerber,et al.  Atomic Force Microscope , 2020, Definitions.

[10]  Z. Shao,et al.  Cryoatomic force microscopy of filamentous actin. , 2000, Biophysical journal.

[11]  Z. Shao,et al.  Promises and problems of biological atomic force microscopy , 1993, Journal of microscopy.

[12]  Z. Shao,et al.  High resolution surface structure of E. coli GroES oligomer by atomic force microscopy , 1996, FEBS letters.

[13]  A. Engel,et al.  Imaging streptavidin 2D crystals on biotinylated lipid monolayers at high resolution with the atomic force microscope , 1999, Journal of microscopy.

[14]  Z. Shao,et al.  High‐resolution atomic‐force microscopy of DNA: the pitch of the double helix , 1995, FEBS letters.

[15]  Z. Shao,et al.  Biological atomic force microscopy: what is achieved and what is needed , 1996 .

[16]  A. Engel,et al.  Voltage and pH-induced channel closure of porin OmpF visualized by atomic force microscopy. , 1999, Journal of molecular biology.

[17]  M. Whittaker,et al.  Molecular structure of F-actin and location of surface binding sites , 1990, Nature.

[18]  Z. Shao,et al.  Staphylococcal alpha-hemolysin can form hexamers in phospholipid bilayers. , 1998, Journal of molecular biology.

[19]  T. Mitchison,et al.  Actin dynamics in vivo. , 1997, Current opinion in cell biology.

[20]  F. Pattus,et al.  Method for forming two-dimensional paracrystals of biological filaments on lipid monolayers. , 1990, Journal of electron microscopy technique.

[21]  H. Butt,et al.  Electrostatic interaction in atomic force microscopy. , 1991, Biophysical journal.

[22]  J. D. Pardee,et al.  [18] Purification of muscle actin , 1982 .

[23]  W. Reeves Royal Microscopical Society , 1873, Nature.

[24]  K A Taylor,et al.  Formation of 2-D paracrystals of F-actin on phospholipid layers mixed with quaternary ammonium surfactants. , 1992, Journal of structural biology.

[25]  W. Kabsch,et al.  Atomic model of the actin filament , 1990, Nature.

[26]  A. Engel,et al.  Mapping flexible protein domains at subnanometer resolution with the atomic force microscope , 1998, FEBS letters.

[27]  J. Spudich,et al.  Purification of muscle actin. , 1982, Methods in enzymology.

[28]  Z. Shao,et al.  Submolecular resolution of single macromolecules with atomic force microscopy , 1998, FEBS letters.

[29]  M. Schmutz,et al.  Two‐dimensional crystallization of proteins on planar lipid films and structure determination by electron crystallography * , 1994, Biology of the cell.

[30]  P. Hansma,et al.  Scanning tunneling microscopy and atomic force microscopy: application to biology and technology. , 1988, Science.