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Ca+ Tropomyosin alpha-1 chain Tropomyosin beta chain Beta-1 adrenergic receptor Beta-2 adrenergic receptor Adenylate cyclase type 9 cAMP-dependent protein kinase catalytic subunit beta cAMP-dependent protein kinase catalytic subunit gamma cAMP-dependent protein kinase type I-alpha regulatory subunit cAMP-dependent protein kinase type I-beta regulatory subunit cAMP-dependent protein kinase type II-alpha regulatory subunit cAMP-dependent protein kinase type II-beta regulatory subunit Myosin light chain kinase, smooth muscle cAMP-dependent protein kinase type I-alpha regulatory subunit Voltage- depenent L-type calcium channel Voltage- depenent L-type calcium channel Voltage- depenent L-type calcium channel Ryanodine receptor 2 Myosin light chain 3 Myosin LC-P Actin, cytoplasmic 1 Sodium/calcium exchanger 1 Voltage-gated T-type calcium channel (ICaT) Voltage-gated T-type calcium channel (ICaT) Voltage- dependent P/Q-type calcium channel subunit alpha-1A Voltage- dependent L-type calcium channel subunit beta-1 Voltage- dependent calcium channel subunit alpha-2/delta-2 Myosin light chain kinase, smooth muscle Serine/threonine- protein phosphatase PP1-beta catalytic subunit Calmodulin-1 Sarcoplasmic/endoplasmic reticulum calcium ATPase 2 Calmodulin-1 cAMP-dependent protein kinase catalytic subunit alpha Timolol Timolol cAMP Ca+ Ca+ Ca+ Ca+ Na+ Na+ Ca+ Ca+ Ca+ Ca+ ATP PPi ATP H2O Ca+ ADP Pi Ca+ Magnesium Magnesium Calcium Manganese Ca+ Ca+ Troponin Troponin G-protein signalling cascade Muscle Contraction Muscle Relaxation Other Intracellular Signaling Pathways Muscle Contraction Cardiac Myocyte Sarcoplasmic Reticulum Calcium binding to troponin displaces tropomyosin from the myosin binding sites on the actin filaments Myosin, with an ADP and phosphate attached, binds to actin to form a cross bridge. Myosin performs a powerstroke, drawing the actin filaments together. Many actin filaments pulled together at the same time leads to muscle contraction. Actin Filament Myosin Filament Single Sarcomere Functioning Unit The beta-1 adrenergic receptor is coupled to and activates the G-protein signalling cascade. The G-protein signalling cascade activates protein kinase which activates calcium channels on the membrane, resulting in calcium influx. Calcium activates the ryanodine receptor on the sarcoplasmic reticulum. Time Membrane potential (mV) -40mV (threshold) Pacemaker Action Potential Phase 0- depolarization Phase 3- repolarization Phase 4-spontaneous depolarization With propranolol Without propranolol Timolol, by antagonizing beta-1 adrenergic receptors, decreases intracellular Ca2+. Phase 4 spontaneous depolarization is Ca2+ dependent, therefore propranolol decreases the slope of phase 4, increasing the time it takes for threshold to be reached and thus slows heart rate Cytosol Since calcium is linked to activation of muscle contraction. Reducing cytosolic calcium levels reduces muscle contraction Timolol enters the bloodstream and once it reaches the heart, it antagonizes the beta-1 adrenergic receptor on the cell membrane of myocytes such that epinephrine stimulation of the heart is reduced. T-tubule Activation of beta-2adrenergic recepotrs activates the Gs signaling cascade leading to increased levels of cAMP and activation of protein kinase A. Timolol activates beta-2-adrenergic receptors in the smooth muscle of the respiratory airways Accumulation of myosin LC leads to increased relaxation of the airway smooth muscles Myosin light chain kinase phosphorylates myosin light chain, whereas myosin light chain phosphatase dephosphorylates myosin LC-P Myosin LC-P interacts with actin to produce smooth muscle contraction. Since there are low levels of myosin LC-P, contraction is inhibited Caclium usually binds to calmodulin to activate myosin light chain kinase. Since Ca2+ levles are low, activation of myosin light chain kinase is reduced. PKA may also phosphorylate Gq-coupled receptors leading to a cascade of intracellular signals which reduce intracellular Ca2+ PKA inactivates myosin light chain kinase by phosphorylating it
Mitochondria Calcium TPM1 TPM2 ADRB1 ADRB2 ADCY9 PRKACB PRKACG PRKAR1A PRKAR1B PRKAR2A PRKAR2B MYLK PRKAR1A CACNA1C CACNA2D2 CACNB1 RYR2 MYL3 MYL3 ACTB SLC8A1 CACNA1H CACNA1G CACNA1A CACNB1 CACNA2D2 MYLK PPP1CB CALM1 ATP2A2 CALM1 PRKACA Timolol Timolol cAMP Calcium Calcium Calcium Calcium Sodium Sodium Calcium Calcium Calcium Calcium Adenosine triphosphate Pyrophosphate Adenosine triphosphate Water Calcium Adenosine diphosphate Phosphate Calcium Calcium Calcium Troponin Troponin G-protein signalling cascade Muscle Contraction Muscle Relaxation Other Intracellular Signaling Pathways Muscle Contraction
Ca+ TPM1 TPM2 ADRB1 ADRB2 ADCY9 PRKACB PRKACG PRKAR1A PRKAR1B PRKAR2A PRKAR2B MYLK PRKAR1A CACNA1C CACNA2D2 CACNB1 RYR2 MYL3 MYL3 ACTB SLC8A1 CACNA1H CACNA1G CACNA1A CACNB1 CACNA2D2 MYLK PPP1CB CALM1 ATP2A2 CALM1 PRKACA Timolol Timolol cAMP Ca+ Ca+ Ca+ Ca+ Na+ Na+ Ca+ Ca+ Ca+ Ca+ ATP PPi ATP H2O Ca+ ADP Pi Ca+ Mg2+ Mg2+ Ca2+ Mn2+ Ca+ Ca+ Troponi Troponi G-p s c Mus Con Mus Rel O I S P Mus Con Cardiac Myocyte Sarcoplasmic Reticulum Calcium binding to troponin displaces tropomyosin from the myosin binding sites on the actin filaments Myosin, with an ADP and phosphate attached, binds to actin to form a cross bridge. Myosin performs a powerstroke, drawing the actin filaments together. Many actin filaments pulled together at the same time leads to muscle contraction. Actin Filament Myosin Filament Single Sarcomere Functioning Unit The beta-1 adrenergic receptor is coupled to and activates the G-protein signalling cascade. The G-protein signalling cascade activates protein kinase which activates calcium channels on the membrane, resulting in calcium influx. Calcium activates the ryanodine receptor on the sarcoplasmic reticulum. Time Membrane potential (mV) -40mV (threshold) Pacemaker Action Potential Phase 0- depolarization Phase 3- repolarization Phase 4-spontaneous depolarization With propranolol Without propranolol Timolol, by antagonizing beta-1 adrenergic receptors, decreases intracellular Ca2+. Phase 4 spontaneous depolarization is Ca2+ dependent, therefore propranolol decreases the slope of phase 4, increasing the time it takes for threshold to be reached and thus slows heart rate Cytosol Since calcium is linked to activation of muscle contraction. Reducing cytosolic calcium levels reduces muscle contraction Timolol enters the bloodstream and once it reaches the heart, it antagonizes the beta-1 adrenergic receptor on the cell membrane of myocytes such that epinephrine stimulation of the heart is reduced. T-tubule Activation of beta-2adrenergic recepotrs activates the Gs signaling cascade leading to increased levels of cAMP and activation of protein kinase A. Timolol activates beta-2-adrenergic receptors in the smooth muscle of the respiratory airways Accumulation of myosin LC leads to increased relaxation of the airway smooth muscles Myosin light chain kinase phosphorylates myosin light chain, whereas myosin light chain phosphatase dephosphorylates myosin LC-P Myosin LC-P interacts with actin to produce smooth muscle contraction. Since there are low levels of myosin LC-P, contraction is inhibited Caclium usually binds to calmodulin to activate myosin light chain kinase. Since Ca2+ levles are low, activation of myosin light chain kinase is reduced. PKA may also phosphorylate Gq-coupled receptors leading to a cascade of intracellular signals which reduce intracellular Ca2+ PKA inactivates myosin light chain kinase by phosphorylating it
Mitochondria Ca2+ TPM1 TPM2 ADRB1 ADRB2 ADCY9 PRKACB PRKACG PRKAR1A PRKAR1B PRKAR2A PRKAR2B MYLK PRKAR1A CACNA1C CACNA2D2 CACNB1 RYR2 MYL3 MYL3 ACTB SLC8A1 CACNA1H CACNA1G CACNA1A CACNB1 CACNA2D2 MYLK PPP1CB CALM1 ATP2A2 CALM1 PRKACA Timolol Timolol cAMP Ca2+ Ca2+ Ca2+ Ca2+ Na+ Na+ Ca2+ Ca2+ Ca2+ Ca2+ ATP Ppi ATP H2O Ca2+ ADP Pi Ca2+ Ca2+ Ca2+ Troponi Troponi G-p s c Mus Con Mus Rel O I S P Mus Con