Interactions of benzodiazepine derivatives with annexins.
Human annexins III and V, members of the annexin family of calcium- and membrane-binding proteins, were complexed within the crystals with BDA452, a new 1,4-benzodiazepine derivative by soaking and co-crystallization methods. The crystal structures of the complexes were analyzed by x-ray crystallography and refined to 2.3- and 3.0-A resolution. BDA452 binds to a cleft which is located close to the N-terminus opposite to the membrane binding side of the proteins. Biophysical studies of the interactions of various benzodiazepine derivatives with annexins were performed to analyze the binding of benzodiazepines to annexins and their effects on the annexin-induced calcium influx into phosphatidylserine/phosphatidylethanolamine liposomes. Different effects were observed with a variety of benzodiazepines and different annexins depending on both the ligand and the protein. Almost opposite effects on annexin function are elicited by BDA250 and diazepam, its 7-chloro-derivative. We conclude that benzodiazepines modulate the calcium influx activity of annexins allosterically by stabilizing or destabilizing the conducting state of peripherally bound annexins in agreement with suggestions by Kaneko (Kaneko, N., Ago, H., Matsuda, R., Inagaki, E., and Miyano, M. (1997) J. Mol. Biol., in press).
Hofmann
A
A
Max-Planck-Institut für Biochemie, Abt. Strukturforschung, Am Klopferspitz 18a, D-82152 Martinsried, Germany.
Escherich
A
A
Lewit-Bentley
A
A
Benz
J
J
Raguenes-Nicol
C
C
Russo-Marie
F
F
Gerke
V
V
Moroder
L
L
Huber
R
R
D000595:Amino Acid Sequence; D000818:Animals; D017318:Annexin A3 / Q000737:chemistry / Q000378:metabolism; D017304:Annexin A5 / Q000737:chemistry / Q000378:metabolism; D017302:Annexins / Q000737:chemistry / Q000378:metabolism*; D014151:Anti-Anxiety Agents / Q000737:chemistry / Q000378:metabolism*; D001569:Benzodiazepines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D018360:Crystallography, X-Ray; D003975:Diazepam / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D008081:Liposomes; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011949:Receptors, Cholecystokinin / Q000037:antagonists & inhibitors; D017386:Sequence Homology, Amino Acid; D013552:Swine
D017304:Annexin A5; D017302:Annexins; D014151:Anti-Anxiety Agents; C110385:BDA 250; C110384:BDA 452; D008081:Liposomes; D011949:Receptors, Cholecystokinin; D001569:Benzodiazepines; D017318:Annexin A3; D003975:Diazepam; D002118:Calcium