The radius of gyration of native and reductively methylated myosin subfragment-1 from neutron scattering.
Reductive methylation of nearly all lysine groups of myosin subfragment-1 (S1) was required for crystallization and solution of its structure at atomic resolution. Possible effects of such methylation on the radius of gyration of chicken skeletal muscle myosin S1 have been investigated by using small-angle neutron scattering. In addition, we have investigated the effect of MgADP.Vi, which is thought to produce an analog of the S1.ADP.Pi state, on the S1 radius of gyration. We find that although methylation of S1, with or without SO42- ion addition, does not significantly alter the structure, addition of ADP plus vanadate does decrease the radius of gyration significantly. The S1 crystal structure predicts a radius of gyration close to that measured here by neutron scattering. These results suggest that the overall shape by crystallography resembles nucleotide-free S1 in solution. In order to estimate the effect of residues missing from the crystal structure, the structure of missing loops was estimated by secondary-structure prediction methods. Calculations using the complete crystal structure show that a simple closure of the nucleotide cleft by a rigid-body torsional rotation of residues (172-180 to 670) around an axis running along the base of the cleft alone does not produce changes as large as seen here and in x-ray scattering results. On the other hand, a rigid body rotation of either the light-chain binding domain (767 to 843 plus light chains) or of a portion of 20-kDa peptide plus this domain (706 to 843 plus light chains) is more readily capable of producing such changes.
Stone
D B
DB
Cardiovascular Research Institute, University of California, San Francisco 94143, USA.
Schneider
D K
DK
Huang
Z
Z
Mendelson
R A
RA
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D008745:Methylation; D008958:Models, Molecular; D018482:Muscle, Skeletal; D015879:Myosin Subfragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D009218:Myosins / Q000737:chemistry / Q000378:metabolism*; D009502:Neutrons; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation
D000596:Amino Acids; D015879:Myosin Subfragments; D009218:Myosins