Actin dynamics studied by solid-state NMR spectroscopy.
Solid-state nuclear magnetic resonance spectroscopy was used to study the motion of 2H and 19F probes attached to the skeletal muscle actin residues Cys-10, Lys-61 and Cys-374. The probe resonances were observed in dried and hydrated G-actin, F-actin and F-actin-myosin subfragment-1 complexes. Restricted motion was exhibited by 19F probes attached to Cys-10 and Cys-374 on actin. The dynamics of probes attached to dry cysteine powder or F-actin were very similar and the binding of myosin had little effect indicating that the local probe environment imposes the major influence on motion in the solid state. Correlation times determined for the solid state probes indicated that they were undergoing some rapid internal motion in both G-actin and F-actin such as domain twisting. The probe size influenced the motion in G-actin and appeared to sense monomer rotation but not in F-actin where segmental mobility and intramonomer co-ordination appeared to dominate.
Phillips
L
L
Department of Anatomy, University of Sydney, N.S.W., Australia.
Separovic
F
F
Cornell
B A
BA
Barden
J A
JA
dos Remedios
C G
CG
D000096:Acetone / Q000031:analogs & derivatives; D000199:Actins / Q000737:chemistry*; D000818:Animals; D003903:Deuterium / Q000737:chemistry; D005461:Fluorine / Q000737:chemistry; D007460:Iodoacetamide / Q000737:chemistry; D017879:Isothiocyanates; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008628:Mercury; D008782:Methylurea Compounds / Q000737:chemistry; D009132:Muscles / Q000166:cytology; D009218:Myosins / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D013861:Thiocyanates / Q000737:chemistry; D005464:Fluorobenzenes; D005466:Fluorocarbons
D000096:Acetone; D000199:Actins; D003903:Deuterium; D005461:Fluorine; D007460:Iodoacetamide; D017879:Isothiocyanates; D046911:Macromolecular Substances; D008628:Mercury; D008782:Methylurea Compounds; D009218:Myosins; D009942:Organometallic Compounds; D013861:Thiocyanates; C002249:O-methylisourea; C003064:pentafluorophenylisothiocyanate; D005464:Fluorobenzenes; D005466:Fluorocarbons; C069522:(trifluoromethyl)mercuric bromide; C004269:bromotrifluoroacetone