1. Calcium-dependent gating of Voltage-gated Ca 2+ channels Regulation by Calmodulin

2. Ca 2+ fluxes  Internal free calcium  Effect Ion fluxes  Gates  Membrane potential Other transport mechanisms Other stimuli Ca 2+ Ions Transduce Signals Ion Channels of Excitable Membranes, 3 rd Edition, 2001

3. Berne & Levy, Cardiovascular Physiology 6th Edition (1992) Ca ++ Ca 2+ current controls the plateau phase of the cardiac action potential Journal of Physiology (2000) 525.2, pp. 285-298

4. Annu. Rev. Cell Dev. Biol. 2000. 16:521–55 Ca 2+ channels control neurotransmitter release CALCIUM CHANNEL Ca 2+

5. Ca 2+ channels regulate gene expression in hippocampal neurons Current Opinion in Neurobiology 1997, 7:419–429 NATURE |VOL 401 | 14 OCTOBER 1999 |703

6. Ca 2+ channel Structure Annu. Rev. Cell Dev. Biol. 2000. 16:521–55

7. Neuron, Vol. 25, 533–535, March, 2000 Family of Voltage-Gated Ca 2+ Channels L-type P/Q-type

8. Circ Res. 2001; 89:944-956. Ca 2+ channel inactivation is relatively slow I Ca,L

9. Ca 2+ -dependent gating of the L-type Ca 2+ Channel

10. SCIENCE, VOL. 202, 15 DECEMBER 1978 The permeating ion affects Ca 2+ channel inactivation: Calcium Dependent Inactivation (CDI)

11. Ba 2+ Ca 2+ -90 +20 mV Voltage Current 100 ms CDI in a heterologous system

12. Conditional open probability analysis (COPA): Ca 2+ entry enhances inactivation SCIENCE, VOL. 250, 21 DECEMBER 1990 Ba 2+ Ca 2+

13. Ba 2+ Ca 2+ Ba 2+ Ca 2+ CDI is greatest at membrane potentials eliciting peak inward Ca 2+ current SCIENCE VOL. 270 1502

14. 100 mM Ba 2+ out Biophysical JournalVolume 66 April 19941051-1060 CDI in L-type channels reconstituted in bilayers

15. Ca 2+ sensing apparatus resides within Ca V 1.2 (  1C) Rapid effects (< 5 msec) CDI in bilayers Minimal effects of Ca 2+ chelators

16. EF-hand Early candidate: EF- hand in C-terminus

17. CE EC EF-hand region from  1C : necessary and sufficient SCIENCEVOL 2701 DECEMBER 19951502

18. Ca 2+ -binding in the EF-hand is not necessary Proc. Natl. Acad. Sci. USA Vol. 94, pp. 2301–2305, March 1997

19. Biophysical Journal Volume 78 April 2000 1906–1920 EF-hand: an essential for CDI independent of Ca 2+ binding

20. Other region(s) in the C- terminus THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 272, No. 6, Issue of February 7, pp. 3560–3566, 1997 Proc. Natl. Acad. Sci. USA Vol. 95, pp. 3287–3294, March 1998

21. IQ motif IQ motif / calmodulin (CaM) binding domain: CaM as the Ca 2+ sensor NATURE |VOL 399 | 13 MAY 1999 | 159

22. IQ motif is the CaM effector site NATURE |VOL 399 | 13 MAY 1999 | 159

23. CaM binds to the IQ motif in the C-tail

24. NC 1477-15921592-1875 4- WT 1551-1660 CaM is constitutively bound to  1C THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 276, No. 33, Issue of August 17, pp. 30794–30802, 2001

25. Comparison of cation binding affinities of Calmodulin EF hands EC 50 values (  M) Ca 2+ Sr 2+ Ba 2+ Effective ionic radius (Å )1.061.211.38 Phosphodiesterase activity via CaM ~2.5~25>1000 Mol Pharmacol 26:75-82, 1984 CDI correlates with the affinity of the divalent ion for CaM

26. Calcium Dependent Inactivation of L-type Ca 2+ Channels Calmodulin is the Ca 2+ sensor CaM is pre-associated with  1C The IQ motif is the effector domain in  1C The EF-hand is a structural, non Ca 2+ - sensing domain

27. SCIENCEVOL 294318 12 OCTOBER 2001 SCIENCEVOL 294333 12 OCTOBER 2001 Gene expression in hippocampal neurons: LTCs and CaM

28. 100 ms Journal of Physiology (1990), 428, pp. 653-671 Frequency-dependent stimulation of L-type Ca 2+ current: Calcium Dependent Facilitation 2.0 Hz

29. Force-Frequency Relationship Treppe Contraction or “Staircase” Effect Pharm. Rev. 15:601 (1963)

30. CDF is Regulated by CaMKII Proc. Natl. Acad. Sci. USA Vol. 91, pp. 9659-9663, September 1994

31. NATURE CELL BIOLOGY | VOL 2 | MARCH 2000 | 173 CaMKII phosphorylation controls CDF

32. JBC 275: 14354, 2000 Ca 2+ /CaM-dependent kinase II (CaMKII): A molecular clock

33. Calcium Dependent Facilitation is also controlled by the IQ Motif Ile  Ala NATURE |VOL 399 | 13 MAY 1999 | 159

34. CaM is also the Ca 2+ sensor for CDF NATURE |VOL 399 | 13 MAY 1999 | 159

35. Calcium Dependent Facilitation of L-type Ca 2+ Channels Calmodulin is the Ca 2+ sensor The IQ motif is the effector domain in  1C Phosphorylation by CaMKII leads to increased P o

36. Ca 2+ -dependent gating of P/Q-type Ca 2+ Channels

37. NATURE |VOL 411 | 24 MAY 2001 |485 CDI and CDF in P/Q channels

38. CaM is the Ca 2+ sensor? NATURE |VOL 411 | 24 MAY 2001 |485

39. Different kinetics: different effector site? NATURE |VOL 411 | 24 MAY 2001 |485 NATURE |VOL 399 | 13 MAY 1999 |155

40. Different kinetics: different Ca 2+ sensor? nature neuroscience volume 5 no 3 march 2002 210

41. Ionic Dependence of I pCa Inactivation is it really CaM? Neuron, Vol. 20, 797–807, April, 1998 Time constantsCa 2+ Sr 2+ Ba 2+  Fast (ms) 43.6 ± 6.448.8 ± 2.988.5 ± 8.6  Slow (ms) 477 ± 51576 ± 49673 ± 46 PDE activity via CaM~2.5~25>1000

42. Calcium Dependent Gating of P/Q Ca 2+ Channels Kinetics of inactivation and facilitation differ from L-type channel gating Calmodulin appears to be the Ca 2+ sensor and the IQ motif is one effector domain in  1A CBD may be another effector domain The Ca 2+ -binding protein CaBP1 regulates inactivation in a Ca 2+ -independent manner