We have used x-ray small-angle solution scattering to investigate the conformation of several intact myosin S1 kinetic intermediates. The structure of a smaller portion of S1 (called S1Dc) complexed with ADP BeFx, ADP AlF4 and ADP Vi analogs has been determined, and we used S1 + analogs and ATP (actively hydrolyzing S1) to selectively populate these intermediates. The results we obtained are correlated with the S1 and S1Dc crystallographic results. We find that there is a unitary large scale conformational change in S1 (which seems to occur just before or during ATP hydrolysis) which acts to cock the acto-myosin system for subsequent energy transduction. Modeling the observed changes in scattering curves using F-actin and S1 structures indicates that the longitudinal projection of the force-producing portion of the working stroke is less than 6 nm. This work has recently been published in the Journal of Molecular Biology. This year we studied more kinetic intermediates, using new ATP analogues.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001209-19
Application #
2788388
Study Section
Project Start
Project End
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
19
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Vickers, Chelsea; Liu, Feng; Abe, Kento et al. (2018) Endo-fucoidan hydrolases from glycoside hydrolase family 107 (GH107) display structural and mechanistic similarities to ?-l-fucosidases from GH29. J Biol Chem 293:18296-18308
Nguyen, Phong T; Lai, Jeffrey Y; Lee, Allen T et al. (2018) Noncanonical role for the binding protein in substrate uptake by the MetNI methionine ATP Binding Cassette (ABC) transporter. Proc Natl Acad Sci U S A 115:E10596-E10604
Aleman, Fernando; Tzarum, Netanel; Kong, Leopold et al. (2018) Immunogenetic and structural analysis of a class of HCV broadly neutralizing antibodies and their precursors. Proc Natl Acad Sci U S A 115:7569-7574
Herrera, Nadia; Maksaev, Grigory; Haswell, Elizabeth S et al. (2018) Elucidating a role for the cytoplasmic domain in the Mycobacterium tuberculosis mechanosensitive channel of large conductance. Sci Rep 8:14566
Lal, Neeraj K; Nagalakshmi, Ugrappa; Hurlburt, Nicholas K et al. (2018) The Receptor-like Cytoplasmic Kinase BIK1 Localizes to the Nucleus and Regulates Defense Hormone Expression during Plant Innate Immunity. Cell Host Microbe 23:485-497.e5
Pluvinage, Benjamin; Grondin, Julie M; Amundsen, Carolyn et al. (2018) Molecular basis of an agarose metabolic pathway acquired by a human intestinal symbiont. Nat Commun 9:1043
Beyerlein, Kenneth R; J├Ânsson, H Olof; Alonso-Mori, Roberto et al. (2018) Ultrafast nonthermal heating of water initiated by an X-ray Free-Electron Laser. Proc Natl Acad Sci U S A 115:5652-5657
Yoshizawa, Takuya; Ali, Rustam; Jiou, Jenny et al. (2018) Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites. Cell 173:693-705.e22
Noach, Ilit; Ficko-Blean, Elizabeth; Pluvinage, Benjamin et al. (2017) Recognition of protein-linked glycans as a determinant of peptidase activity. Proc Natl Acad Sci U S A 114:E679-E688
Robb, Melissa; Hobbs, Joanne K; Woodiga, Shireen A et al. (2017) Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. PLoS Pathog 13:e1006090

Showing the most recent 10 out of 604 publications