Consequences of placing an intramolecular crosslink in myosin S1.
This paper describes the placement of a crosslinking agent (dibromobimane) between two thiols (Cys-522 and Cys-707) of a fragment, "S1," of the motor protein, myosin. It turns out that fastening the first anchor of the crosslinker is easy and rapid, but fastening the second anchor (Cys-522) is very temperature dependent, taking 30 min at room temperature but about a week on ice. Moreover, crystallography taken at 4 degrees C would seem to predict that the linkage is impossible, because the span of the crosslinking agent is much less than the interthiol distance. The simplest resolution of this seeming paradox is that structural fluctuations of the protein render the linkage increasingly likely as the temperature increases. Also, measurements of the affinity of MgADP for the protein, as well as the magnetic resonance of the P-atoms of the ADP once emplaced, suggest that binding the first reagent anchor to Cys-707 initiates an influence that travels to the rather distant ADP-binding site, and it is speculated what this "path of influence" might be.
Konno
K
K
Hokkaido University, Hakodate, Japan.
Ue
K
K
Khoroshev
M
M
Martinez
H
H
Ray
B
B
Morales
M F
MF
D000244:Adenosine Diphosphate / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D001643:Bridged Bicyclo Compounds / Q000737:chemistry; D003198:Computer Simulation; D003432:Cross-Linking Reagents / Q000737:chemistry; D005453:Fluorescence; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D015879:Myosin Subfragments / Q000737:chemistry*; D011487:Protein Conformation; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013696:Temperature
D001643:Bridged Bicyclo Compounds; D003432:Cross-Linking Reagents; D015879:Myosin Subfragments; D013438:Sulfhydryl Compounds; D000244:Adenosine Diphosphate; C037094:dibromobimane