Test 2 Flashcards
What are some of the uses for motor proteins we discuss?
Organelle migration, motion of flagella, movement of some proteins along DNA
Describe the steps of myosin/actin binding
- Resting State
- Myosin heads are “cocked” and attached to ADP and an inorganic phosphate (Pi), but they are not yet bound to actin.
- Tropomyosin is blocking the active sites on actin, preventing myosin from binding to actin. This is controlled by troponin, which binds calcium ions when released from the sarcoplasmic reticulum (SR). - Calcium Ion Release
- When an action potential reaches the muscle, calcium ions are released from the sarcoplasmic reticulum into the cytoplasm.
-Calcium binds to troponin, causing a conformational change in the troponin-tropomyosin complex. - Exposure of Binding Sites
- The conformational change in the troponin-tropomyosin complex shifts tropomyosin away from the myosin-binding sites on the actin filament.
This exposes the binding sites on actin, allowing myosin heads to bind to actin. - Cross-Bridge Formation
- The myosin heads, which have ADP and Pi bound, now attach to the exposed binding sites on actin. This forms the cross-bridge between myosin and actin. - Power Stroke
-Once the myosin head binds to actin, the release of Pi from myosin causes a conformational change in the myosin head, pulling the actin filament toward the center of the sarcomere. This is the power stroke.
-This action generates force, causing the sliding of the actin filament relative to the myosin filament, which leads to muscle contraction. - ADP Release
-After the power stroke, ADP is released from the myosin head. - ATP Binding and Myosin Head Detachment
-A new molecule of ATP binds to the myosin head, causing the myosin head to detach from the actin filament.
-The ATP is hydrolyzed into ADP and Pi, which re-cocks the myosin head, readying it for another cycle of binding and pulling.
Describe the structural details of myosin
- Six subunits: 2 heavy and 4 light chains
- Fibrous and globular domains
- Aggregate to form thick filaments
- Cleavage by trypsin and papain
- Trypsin cleaves the light fibrous
tail from the globular heads
and then the two heads are
cleaved from each other by
papain
Describe how actin comes into play
Actin is involved in muscle contraction.
- Monomeric actin, G-actin (G for globular), associates to form F-actin (F for filamentous)
-F-actin assembly is driven by ATP hydrolysis
-F-actin along with troponin and tropomyosin comprise the thin filament
-1:1 ratio between actin monomers and myosin head groups
-Muscle contraction occurs from thick filaments sliding along thin filaments
How do tropomyosin and troponin regulate the nerve impulses related to muscle contraction
Troponin and tropomyosin are in complex
- Troponin is a Ca2+ binding protein
- The nerve impulse causes Ca2+ release and subsequent structural changes, exposing myosin binding sites
What do enzymes do?
Catalyze reactions
Enzymes are central to every biochemical process
How do enzymes bind substrate?
Through geometric and electronic complementarity
Name the seven types of enzymes and what they do
Classified according to reaction type:
- Lyases: Addition of groups to double bonds or formation of double bonds by removal of groups
- Isomerases: Transfer of groups within molecules to produce isomeric forms
- Ligases: Formation of bonds by condensation reactions coupled to ATP cleavage
- Hydrolases: Hydrolysis reactions: Transfer of functional groups to H2O
- Oxidoreductases: Transfer of electrons
- Transferases: Group transfer reactions
- Translocases: Facilitate the movement of molecules, such as proteins or other substances, across cellular membranes
What is the enzyme suffix?
-ase
Give the scheme for a simple enzyme catalyzed rxn which converts a single substrate into a single product.
E+S <–> ES <–> EP <–> E+P
Do enzymes affect the rxn equilibria?
No. They just lower the activation energy, thereby speeding up the reaction
- Accelerate interconversion of S and P. The free energy used to lower activation energy comes from binding energy
What does the equilibrium constant Keq describe?
The equilibrium between S <–>P
K’eq= [P]/[S]
When is binding optimal between E and S?
In the transition state.
- Weak binding interactions between E and S provide a substantial driving force for catalysis, creating net lower activation energy
- Binding energy also gives specificity
What does the Rate of product formation depend on?
Activation energy
Define Km
Km: Substrate concentration at which an enzyme-catalyzed reaction proceeds at ½ its maximum velocity
Define V0
Vo: Initial rate or velocity of an enzyme reaction during which [S] does not change drastically
Define Vmax
Maximum velocity of an enzyme reaction when the binding site is saturated with substrate
Define Kcat
kcat: Number of substrate molecules converted to product in a given amount of time on a single enzyme molecule (turnover number)
Complex function of several rate constants - dependent on the rate limiting step!
What are the types of reversible enzyme inhibition?
Competitive
Uncompetitive
Noncompetitive (mixed)
What do enzyme inhibitors do?
- Reduce enzyme efficiency
- Can be good drugs if they inhibit
essential (viral or bacterial)
enzymes - Reversible or irreversible
Define Competitive Inhibition
- Inhibitor binds in the enzyme active
site – competes with normal
substrate binding (forms EI
complex) - Prevents substrate from binding
and reacting - Usually looks like substrate (or
transition state of reaction!)
Describe competitive inhibitions effect on kinetics
Vmax remains constant
Km increases
- Dependent on alpha
Apparent Km: aKm
Define uncompetitive inhibition
Inhibitor binds to enzyme-substrate complex at site distinct from active site
Distorts the active site making it inactive via conformational change
Describe uncompetitive inhibition effect on kinetics
Vmax & Km decrease
Both dependent on a’
Apparent Km: Km/a’
Apparent Vmax: Vmax/a’