EXAM 3 Flashcards
skeletal muscle is responsible for _
- 400+ in body
- ~40-45% of body weight
all movement and support
skeletal muscle is mostly made up of _
- 20% protein
- 5% other: fats, CHO, high energy phosphates, urea, lactic acid, enzymes, Na+, K+, Cl-, and minerals - Ca2+, Mg, and phosphorous
water (75%)
skeletal muscle is _ in appearance
- structure is fascia and connective tissue sheaths which separate individual muscles and hold muscles in place
- under fascia: epimysium
- under epimysium: fascicles
- there are ~ 150 fibers/fasciculus
striated
connective tissue surrounding muscle
epimysium
- bundle of muscle cells (fibers) surrounded by perimysium
- each fiber in _ is surrounded by endomysium
- under endomysium is _, cell membrane
- connective tissue surrounds muscle and forms a network that extends throughout the muscle
- fasciulus
- sarcolemma
bundles of muscle fibers enclosed by perimysium
fascicles
- cytoplasm of muscle cell
- under sarcolemma
- contains contractile proteins, enzymes, fat, and glycogen particles, nuclei, and specialized cellular organelles
sarcoplasm
greatest amounts:
- myosin
- actin
- tropomyosin
- also, large amounts of myoglobin
proteins in skeletal muscle
muscles cells are: _ and _
- multi-nucleated
- striated appearance
- embedded in sarcoplasm
- network of channels and vesicles
- lies parallel to the myofibrils
- lateral end: sac-like cisternae which stores Ca2+
sarcoplasmic reticulum (SR)
- perpendicular to myofibril
- _ and two terminal cisternae are the region of the Z line
- called a triad
- two triads/sacromere
- one sacromere: Z line to Z line
- Tubules open to the outside of each muscle fiber
- functioning as a network, spreading the AP from outer to inner portions of muscle fiber
Transverse Tubules (T-Tubules)
- composed of 6 polypeptide chains
- 2 heavy
- 4 light
- one myosin filament is made up of 200 or more myosin molecules
- exactly 1.6 micrometers in length
myosin molecule
Myosin molecule:
- _ are in center
- _ coming out from center (0.2 micrometers)
- hinges at two points
- arm separates from the filament
- where the head attaches to the arm
- filament is twisted, cross-bridges are displaced from previous set by 120 degrees
- ensures that cross-bridge extend in _ from the filament
- tails
- heads
- all directions
- primary protein
- forms backbone of filament
- approx. 3,000 actin/myofibril
- Troponin and Tropomyosin
- two other proteins in filament
actin
Two shapes of actin:
- globular (G-actin)
- Fibrous (F-actin)
Type of shape of actin:
- polymerizes and unfolds
globular (G-actin)
Type of shape of actin:
- double stranded in helix, complete revolution every 70 nanometers
fibrous (F-actin)
- 13 _ molecules in each revolution of the helical strand
- Each G-actin has an _ molecule attached to it
- ADP are purported to be the active sites on the actin for _ interaction
- active sites are staggered, one site on total filament every 2.7 nanometers
- each actin is 1mm long
- G-actin
- ADP
- cross-bridge
- inserted into Z discs, other end protrudes into sacromere
- in spaces between myosin molecules
filament bases
- actin filaments extending from either side
- into neighboring sacromeres
- passes from myofibril to myofibril
- attaching the myofilaments to each other across the muscle fiber
Z line
Basic contractile unit, Z line to Z line
sacromere
- associated with the actin filament
- loosely connected to the F-actin strands
- wrap themselves spirally around the sides of the F-actin helix
- at rest, _ molecules lie on top of the actin sites, inhibiting interaction between actin and myosin
- each _ covers about 7 active sites
tropomyosin
- protein attached near one end of each tropomyosin molecule
- complex of 3 protein subunits
- _ complex may attach the tropomyosin to the actin
- each subunit plays a role in the contractile process
troponin
Troponin complex:
- strong affinity for actin
Troponin I (TnI)
Troponin complex:
- strong affinity for tropomyosin
Troponin T (TnT)
Troponin complex:
- strong affinity for calcium
Troponin C (TnC)
Troponin complex includes 3 subunits:
- TnI
- TnT
- TnC
- alternating light and dark bands along the length of the muscle fiber gives its characteristic striated appearance
- lighter area: I band
- darker area: A band
sacromere ultrastructure
sacromere ultrastructre:
- when light passes through this band, its velocity is the same
I band (isotropic)
sacromere ultrastructure:
- light does not scatter equally
A band (anisotropic)
sacromere ultrastructure:
- Z line bisects the _ and adheres to the sarcolemma, adding stability to the sacromere
I band
sacromere ultrastructure:
- Z line has _ (maintains spacing of actin) and _ (connects Z lines of different myofibrils together)
- position of the actin and myosin redults in an overlap of the filaments in the sacromere
- alpha actin
- desmin
sacromere ultrastructure:
- center of the _ is the H band
- lighter area due to the absence of the actin filaments in the region
A band
sacromere ultrastructure:
- central part of _ is bisected by the M line
H band
sacromere ultrastructure:
- _ is protein structures that support the arrangement of the myosin filaments
- also has myomesin which provides an anchor for titin (elastic filament)
- helps maintain centering between _ and _
- CK, provides ATP from CP
- M line
- Z line, M line
- Tropomyosin and troponin regulate the interaction between actin and myosin proteins of thick and thin filaments
- During contraction, cross-bridges attach between actin and myosin
- Two filaments slide over each other when energy is provided by the hydrolysis of ATP
Sliding Filament Theory (AF Huxley & Neidergerke, 1954)
- skeletal muscle contracts only after stimulation from a motor neuron
- normally, each motor neuron branches several times and stimulates a few to several hundred muscle fibers
neuromuscular bases of contraction
- motor neuron (cell, etc.)
- muscle fibers it innervates
- at muscle site: neuromuscular junction, motor and end plate, myoneural junction
motor unit
- initiation of the action potential by the motor neuron
- transmission of the AP across the motor end plate to the muscle fiber
contraction begins
~ 200-300 vesicles of acetylcholine (ACH, neurotransmitter) are released into the gap between the motor neuron and the motor end plate (cleft)
Action potential reaches NMJ
- reaction causes an increases in permeability to sodium ions, resulting in depolarization of the sarcolemma or end-plate potential
- if end-plate potential is large enough to exceed a threshold (depending on skeletal muscle type), the nerve impulse will be successfully transformed into a muscle impulse
- the impulse travels in all directions over the muscle membrane when being transmitted
- deep into the fiber through the transverse tubules (T-tubules)
ACH diffused across the gap and reacts with receptor molecules in the sarcolemma
- as the AP is transmitted throughout the fiber
- the membranes of the cisterane in the SR becomes more permeable to Ca++
- Ca2+ diffuses into the sarcoplasm of the fiber
- once the Ca 2+ concentration is high enough, (100x increase), the Ca2+ binds with the TnC molecule
- Binding of Ca2+ to the TnC causes a propositional change of the Tn, which also effects the positioning of the tropomyosin, moving it deeper into the groove between the two actin strands
wave of depolarization reaches T tubule
- two different iso forms
- fast muscle
- slow muscle
TnC
- fast contain low binding sites for Ca2+
- site I & site II
- slow have only one binding site
- both sites must be filled to trigger contraction
TnC
- there is a _ with binding that exposes a hydrophobic cavity (the TnI binding site)
- alters the interaction between TnI and TnC
- instead of TnI binding to actin, it perferentially switches to binding domain on TnC, allowing actin and myosin to interact
conformational change
- _ skeletal muscle has no site I
- _ and _ are activated by one, not two calcium ions by the TnC isoforms subunit
- therefore,
- contraction frequency
- power output
- strength are typically down regulated
- slow
- slow and cardiac muscle
- ubiquitous in eukaryotes
- responsible for a wide range of functions requiring directed movement of molecules, subcellular components, cells, tissues, and whole organisms
- utilize the energy of hydrolysis of ATP
motor proteins
- ATP utilizing motor protein
- generates movement by interaction with actin filaments
- all members of the myosin family share the same basic structure
- a single polypeptide chain: the heavy chain is folded to form the head and tail domain
- light chain monomers bind to heavy chain just below the head domain to form the neck domain
- the basic myosin structure is modified to generate the _
myosin
- myosin family of motor proteins
- actin composed of long polymers of the actin monomer subunits
- ubiquitous in animal and plant cells
- major component of the cytoskeleton
- from the ‘ thin filaments ; in skeletal and cardiac muscle
myosin generates movement by interaction with actin filaments
- muscle is comprised of interdigititating filaments of actin and myosin polymers
- during muscle contraction, myosin heads move over and bind to an adjacent actin filament (swinging cross-bridge hypothesis)
- hydrolysis of ATP in myosin head results in a conformational change in the myosin which cause the neck to wing, pulling on the actin filament
- the net result of many actin=myosin interactions is that the filaments slide, relative to one another, causing the muscle fiber to shorten (sliding filament hypothesis)
skeletal muscle myosin II
- light microscopy
- electron microscopy
- X-ray diffraction
muscle ultrastructure
- resolution is limited by the wavelength of the illuminating light
- for visible light microscopy this limits resolution to approx. o.2 nm
light microscopy
- to improve resolution, we need to use shorter wavelengths
- the de Brogile wavelength is given by: wavelength=h/mv
- to achieve a wavelength of 1 nm, we need to accelerate the electrons to about 100,00 ms-1
- a high voltage is required (100,00v)
- circular magnetic lenses focus electrons onto the sample
- advantages:
- much high resolution (to 1 nm)
- disadvantages:
- samples are dead ( drying, heavy metal staining, vacuum)
- artifacts due to staining procedures are common
electron microscopy
- can achieve resolution below 0.1 nm
- much greater resolution than light or electron microscopy
- fiber diffraction methods can detect changes in the structure of living muscle fibers but:
- can only be used on relatively large arrays of molecules (fiber or protein crystal)
- crystals of large proteins (like myosin) can be extremely difficult to grow
- x-ray crystallography of proteins provides a static picture of a single, possible confirmation of a protein
x- ray diffraction
- optical tweezers
- in-vito motility
- pre-steady state kinetics
myosin dynamics
Myosin dynamics:
- individual myosin heads are attached to surface
- the motion of flourecently-labeled actin filaments, propelled by the myosin heads, can be monitored
in-vito motility
fluorescence techniques:
- _: tryptophan residue fluorescence is sensitive to changes in polarity of its environment
intrinsic fluorescence
- intrinsic fluorescence
- fluorescent substrate (Ex: mant-ATP)
- fluorophore labeling on cysteine residues (Ex: pyrene labeled actin)
- FRET (fluorescence resonance energy transfer) techniques: distance measurements between two fluorophores
- quantum dot: large, intense fluorophore, enables detection of single molecule fluorescence
- chimeric constructs with GFP, YFP
fluorescence techniques
a detailed reaction mechanism for the actin: nucleotide interaction can be determined using _
pre- steady state kinetic techniques
structural studies using _ and _ techniques have been used to study the conformational changes in actin and myosin during muscle contraction
EM and X-ray diffraction
_ techniques measure force generation and step size in myosin heads
single molecule (optical tweezers, in-vito motility)
- contraction cycle of myosin cross-bridges of a muscle shortens a muscle by 1% of its resting length
- consequently, the contraction cycle must be repeated over and over to significantly shorten the whole muscle
single contraction cycle
- when a new ATP attaches to a myosin head, the cross-bridge can detach from the actin
- greater amount of work performed by the muscle
- greater amount of ATP which is cleaved
Fenn Effect
- at the same time contraction is occurring, ACH that stimulated the contraction is being _ by the action of _ (enzyme present at the myoneural junction within the membranes of the motor end plate)
- rapid removal of ACH insures that a single nerve impulse will not cause a continued stimulation of the muscle
- rapidly decomposed
- cholinesterase
- usual duration of an impulse to skeletal muscle is about 20 milliseconds
- in order for contraction to continue:
- continual stimulation of the muscle fiber
impulse duration
the signal to stop contraction is the absence of a nerve impulse at the junction (see notes for diagram)
tetanus
Action Potential stops:
- continually _ located in the walls of the SR pumps the calcium ions out of the _
- active calcium pump
- sarcoplasm
Action Potential stops:
- back into the SR via the _ , and then the calcium diffuses back into the _
- fenestrated collar
- cisternae
Action Potential stops:
- calcium diffuses back into cisternae
- this lowers the concentration of calcium, removing it from the conformation and the _
- fiber returns to its relaxed position
active sites are covered
- actin and myosin uncoupled
- calcium stored in SR
rest
- nerve impulse generated
- ACH released from the vesicles
- sarcolemma depolarized
- muscle impulse transmitted through the fiber
- calcium released from cisternae
- calcium binds to troponin
- actin binding sites activated
- myosin ATPase activated
excitation
- myosin cross-bridges swivel
- release Pi + ADP
- Actin slides over myosin
contraction
- ATP attaches to myosin
- actin and myosin dissociate
- ATP – ADP + Pi
- contraction process repeats
regeneration