Structural determinants of high-affinity binding of ryanoids to the vertebrate skeletal muscle ryanodine receptor: a comparative molecular field analysis.
Ryanodine binds to specific membrane proteins, altering the calcium permeability of intracellular membranes. In this study 19 ryanoids were isolated or synthesized and the structures correlated to the strength of binding to vertebrate skeletal muscle ryanodine receptors. Global minima were determined by employment of molecular mechanics and dynamics augmented by systematic searching of conformational space. Overall, steric and electrostatic factors contribute about equally to the differences in the experimentally determined dissociation constants. The dominant electrostatic interaction is localized to a hydroxyl group in an apolar region of the molecule. The pyrrole and isopropyl groups located together at one pole of the molecule have the greatest effect on steric interactions between ligand and receptor. We suggest ryanodine binds to the receptor with the pyrrole and isopropyl groups buried deep inside a cleft in the protein. This arrangement places special importance on the conformation of the pyrrole and isopropyl groups. In contrast, the opposite pole appears to be positioned at the entrance of the binding pocket because bulky adducts placed in the 9 position of ryanodine alter binding minimally. For example, a fluorescent ryanodine adduct was synthesized which has a dissociation constant close to that of ryanodine. Detailed examination reveals subtle interactions between ryanoid and receptor. In many cases, the major factors altering the strength of binding were found to be conformational alterations in the molecule remote from the site of covalent modification.
Welch
W
W
Department of Biochemistry, University of Nevada, Reno 89557.
Ahmad
S
S
Airey
J A
JA
Gerzon
K
K
Humerickhouse
R A
RA
Besch
H R
HR
Ruest
L
L
Deslongchamps
P
P
Sutko
J L
JL
D000818:Animals; D015220:Calcium Channels / Q000378:metabolism*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002645:Chickens; D004563:Electrochemistry; D008958:Models, Molecular; D008968:Molecular Conformation; D015394:Molecular Structure; D009124:Muscle Proteins / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism*; D011817:Rabbits; D012433:Ryanodine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D015220:Calcium Channels; D009124:Muscle Proteins; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine