Effects of calcium and nucleotides on the structure of insect flight muscle thin filaments.
The structure of the insect flight muscle thin filament has been studied using a Drosophila mutant (Ifm(2)2) which does not contain thick filaments. Thin filaments that are biochemically identical to those of the wild type can be isolated free from thick filament contamination. We show that isolated thin filaments have different symmetries depending upon the calcium concentration. While the filaments mainly contain 13 subunits in six turns of the 5.9 nm genetic helix in the absence of calcium, 50% of the filaments have 28 subunits in 13 turns of the genetic helix at calcium concentrations equivalent to those present during muscle contraction. We also show that the structure (mainly the helical order) of the thin filaments depends on the nature of the nucleotide bound to the actin monomers. Three-dimensional reconstructions of the thin filaments in the presence and absence of calcium show that tropomyosin moves between two different positions on the actin filament. However, in Drosophila the amplitude of the movement as well as the disorder in the positions of the components (tropomyosin, troponin complex) are larger than those generally observed in other species.
Ruiz
T
T
Centre de Genetique Moleculaire, Centre National de la Recherche Scientifique, Gif sur Yvette, France.
Bullard
B
B
Lepault
J
J
D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000648:ultrastructure; D000199:Actins / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics*; D008854:Microscopy, Electron; D009132:Muscles / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D009154:Mutation; D009218:Myosins / Q000032:analysis; D009711:Nucleotides / Q000494:pharmacology*; D011487:Protein Conformation; D012518:Sarcomeres / Q000737:chemistry / Q000201:enzymology; D014335:Tropomyosin / Q000032:analysis
D000199:Actins; D009711:Nucleotides; D014335:Tropomyosin; D009218:Myosins; D002118:Calcium