lecture 5-6 Flashcards

1
Q

Extracellular matrix

A
  1. collagen fibers: very elastic and very strong, create tendons, bone with calcium

The extracellular matrix (ECM) is secreted by cells and surrounds them in tissues. It has long been understood to be the structural support for cells since its characteristics set the characteristics of the tissue (i.e. bone compared to cartilage compared to brain)1.

function:

The extracellular matrix helps cells to bind together and regulates a number of cellular functions, such as adhesion, migration, proliferation, and differentiation. It is formed by macromolecules, locally secreted by resident cells.

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2
Q

Phospholipid bilayer

A
  1. Inner and outer layer are independent of each other
  2. Temperature influences phospholipid bilayer (semi-permeable)
  3. You can regulate amount of rna induced for phospholipid bilayer
  4. Phospholipids are synthesized regularly, by switching phospholipids and replacing they become more saturated
  5. Protieins maintaian position in bilayer via polar and non polar elements
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3
Q

Plasma Membrane

  • Selectively permeable
  • Allows for uptake of key nutrients and elimination of waste products
  • Maintains protected environment for cellular processes
  • Development of internal membranes = increase in complexity
A

Plasma Membrane

  • Selectively permeable
  • Allows for uptake of key nutrients and elimination of waste products
  • Maintains protected environment for cellular processes
  • Development of internal membranes = increase in complexity
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4
Q

Membrane Consists of Proteins in a fluid of lipid molecules

describe bilayer and membrane fluid mosaic model

A

Fluid mosaic model
- Bilayer is not solid, fluid, consistency of olive oil
- Bilayer: lipid molecules existing in a double layer less than 10 nm thick
- Lipid molecules of bilayer vibrate, flex back and forth
- Rarely lipid molecule switch layers
- Exchanges between layers occurs millions of times per second = synthesis
o Lipid molecules and membrane dynamic

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5
Q
  • Membrane function = critically maintaining membrane in fluid state
A
  • Membrane function = critically maintaining membrane in fluid state
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6
Q
  • Mosaic aspect = most membranes contain an assortment of types of proteins
    o Includes proteins in transport, electron transport etc.
    o Larger than lipid molecules, proteins move slower in fluid environment of membrane, small umber of membrane proteins anchor cytoskeleton filaments to membrane and do not move
    o Glycolipids and glycoproteins = carbohydrate groups linked with proteins
A
  • Mosaic aspect = most membranes contain an assortment of types of proteins
    o Includes proteins in transport, electron transport etc.
    o Larger than lipid molecules, proteins move slower in fluid environment of membrane, small umber of membrane proteins anchor cytoskeleton filaments to membrane and do not move
    o Glycolipids and glycoproteins = carbohydrate groups linked with proteins
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7
Q
  • Relative proportions of lipid and protein in membrane vary depending on membrane
  • Plasma membrane = equal amount of protein to lipid
A
  • Relative proportions of lipid and protein in membrane vary depending on membrane
  • Plasma membrane = equal amount of protein to lipid
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8
Q
  • Myelin
    o Sublayer functioning to insulate nerve fibers as cell membranes are composed of mostly lipids (maintaining equilibrium)
A
  • Myelin
    o Sublayer functioning to insulate nerve fibers as cell membranes are composed of mostly lipids (maintaining equilibrium)
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9
Q
  • ## Proteins and other components forming half of bilayer are different from those forming other half
A
  • ## Proteins and other components forming half of bilayer are different from those forming other half
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10
Q

-
- Membrane asymmetry, reflects differences diff. in functions performed on either side of membrane
o Glycolipids + carbs attached to proteins on external side of plasma membrane
o Cytoskeleton binds to proteins on internal side

A
  • Membrane asymmetry, reflects differences diff. in functions performed on either side of membrane
    o Glycolipids + carbs attached to proteins on external side of plasma membrane
    o Cytoskeleton binds to proteins on internal side
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11
Q
  • Hormones and growth binds to receptor proteins found in only externals surface of plasma membrane
A
  • Hormones and growth binds to receptor proteins found in only externals surface of plasma membrane
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12
Q
  • Membrane asymmetry:

o Two sides are distincltly different (vary in shape, size, and numbers)

A
  • Membrane asymmetry:

o Two sides are distincltly different (vary in shape, size, and numbers)

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13
Q

Lipid Fabric of Membrane

  • Lipid moelcuels make up underlying fabric of all biological membranes
  • Collectlively lipid referes to divers group of water insoluble molecules includes fats phospholipids (dominant lipids oin membrane) and steroids
  • Many organisms can self regulate membranes = not too rigid or over fluid
A

Lipid Fabric of Membrane

  • Lipid moelcuels make up underlying fabric of all biological membranes
  • Collectlively lipid referes to divers group of water insoluble molecules includes fats phospholipids (dominant lipids oin membrane) and steroids
  • Many organisms can self regulate membranes = not too rigid or over fluid
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14
Q
  • Phospholipid molecule consisting of head group attached to two chains of C and H = fatty acid
  • Head group = glycerol linked to alcohol or amino acids via phosphate group
  • Property critical to structure and function = amphipathic (both water loving and water hating)
    o Hydrophobic head and hydrophilic tails
    o Polar molecules = hydrophilic and non plar = hydrophobic
A
  • Phospholipid molecule consisting of head group attached to two chains of C and H = fatty acid
  • Head group = glycerol linked to alcohol or amino acids via phosphate group
  • Property critical to structure and function = amphipathic (both water loving and water hating)
    o Hydrophobic head and hydrophilic tails
    o Polar molecules = hydrophilic and non plar = hydrophobic
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15
Q
  • Phospholipids range in degree of unsaturation of fatty acids
    o Straight fatty acid chain = fully saturated, carbons bound to max # atoms
    o Kinked fatty acid chain = Carbon double bond = unsaturated
    o When added to water phoispholipids self assemble 3 types
     Micelle
     Lyposome
     Phos. Bilayer
    o Depends mostly on phos. Concentration
A
  • Phospholipids range in degree of unsaturation of fatty acids
    o Straight fatty acid chain = fully saturated, carbons bound to max # atoms
    o Kinked fatty acid chain = Carbon double bond = unsaturated
    o When added to water phoispholipids self assemble 3 types
     Micelle
     Lyposome
     Phos. Bilayer
    o Depends mostly on phos. Concentration
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16
Q

o Phos. Spontaneously form in these aqueous environ. Bc of hydrophobic effect
 HE: tendency of polar molecule to exclude hydrophobic molecules like fatty acids = aggregation of lipid molecules in structures where fatty acid tails interact with each other
 Polar head groups associate with water
 These arrangements favoured= lowest energy state more likely to occur over any other arrangement

A

o Phos. Spontaneously form in these aqueous environ. Bc of hydrophobic effect
 HE: tendency of polar molecule to exclude hydrophobic molecules like fatty acids = aggregation of lipid molecules in structures where fatty acid tails interact with each other
 Polar head groups associate with water
 These arrangements favoured= lowest energy state more likely to occur over any other arrangement

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17
Q

Fatty acid composition and temp vs membrane fluidity
- Fluidity of lipid bilayer influenced by:
o Type of fatty acids
o Temp.

A

Fatty acid composition and temp vs membrane fluidity
- Fluidity of lipid bilayer influenced by:
o Type of fatty acids
o Temp.

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18
Q
  • Saturated fatty acids = linear allow for compact

- Unsaturated fatty acids = kinks less room for compact

A
  • Saturated fatty acids = linear allow for compact

- Unsaturated fatty acids = kinks less room for compact

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19
Q
  • Temp. affects membrane fluidity
    o Lower temp = lower molecule movement, phospholipid forms semi solid, decrease membrane permeability, inhibit protein complexes within bilayer, proteins cannot change shape due to rigidity
    o Higher temp = increased molecule movement too fluid resulting in loss of integrity, membrane is leaky, ion concentration permeability increases lead to cell death
    o Membrane systems equilibrium = mixed of saturated and unsaturated
A
  • Temp. affects membrane fluidity
    o Lower temp = lower molecule movement, phospholipid forms semi solid, decrease membrane permeability, inhibit protein complexes within bilayer, proteins cannot change shape due to rigidity
    o Higher temp = increased molecule movement too fluid resulting in loss of integrity, membrane is leaky, ion concentration permeability increases lead to cell death
    o Membrane systems equilibrium = mixed of saturated and unsaturated
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20
Q

c organism can adjust fatty acid composition
- Optimal state of fluidity
o Organism self regulate via unsat. and sat. fatty acid concentration regulation proportion

A

hh

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21
Q
  • All fatty acids are initially synthesized as fully sat. molecules w out double C bonds
    o Desaturases, acts on saturated fatty acids by catalyzing and introducing double C
A

hh

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22
Q
  • Sterols:
    o Influences membrane fluidity
    o Cholesterol, found in membrane of animals but not plants or prokaryotes
    o Act as membrane buffers at high temp they restrain movement, low temp. sterols disrupt fatty acids from associating by occupying space
A
  • Sterols:
    o Influences membrane fluidity
    o Cholesterol, found in membrane of animals but not plants or prokaryotes
    o Act as membrane buffers at high temp they restrain movement, low temp. sterols disrupt fatty acids from associating by occupying space
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23
Q

Membrane proteins – a and b + Key functions
- Proteins associated with membrane determine function
- 4 major functional categories:
 2 major types of proteins

A

Membrane proteins – a and b + Key functions
- Proteins associated with membrane determine function
- 4 major functional categories:
 2 major types of proteins

24
Q

o Integral: proteins embedded in phospholipid bilayer = transmembrane proteins bc they interact with both sides of membrane
 Transport proteins = provides channel providing movement of specific molecule membrane proteins = changes shape to shuttle specific molecule from one side of membrane to another (channel protein)
 Enzymatic activity
 Signal transduction= membranes contain receptor proteins binding to specific chemicals like hormones, trigger changes on inner surface of membrane
 Attachment and recognition proteins = proteins exposed to inner and outer parts of membrane are attachment point for range of cytoskeleton elements
• Integral membrane proteins interact with membrane hydrophobic core
• Transmembrane proteins have 2 domains
o Domain interacting with lipid bilayer consists of non-polar amino acids
 Creates channels “tampons” aka alpha helix (secondary structure)
o Domain exposed of either side of membrane composed of polar amino acids

A

o Integral: proteins embedded in phospholipid bilayer = transmembrane proteins bc they interact with both sides of membrane
 Transport proteins = provides channel providing movement of specific molecule membrane proteins = changes shape to shuttle specific molecule from one side of membrane to another (channel protein)
 Enzymatic activity
 Signal transduction= membranes contain receptor proteins binding to specific chemicals like hormones, trigger changes on inner surface of membrane
 Attachment and recognition proteins = proteins exposed to inner and outer parts of membrane are attachment point for range of cytoskeleton elements
• Integral membrane proteins interact with membrane hydrophobic core
• Transmembrane proteins have 2 domains
o Domain interacting with lipid bilayer consists of non-polar amino acids
 Creates channels “tampons” aka alpha helix (secondary structure)
o Domain exposed of either side of membrane composed of polar amino acids

25
Q
  • Peripheral: interact with hydrophilic surface
    o Positioned on surface of membrane do not interact hydrophobic core
    o Found on cytoplasmic side of membrane and form part of cytoskeleton
    o Mad of of mixture of polar and non-polar amino acids
A
  • Peripheral: interact with hydrophilic surface
    o Positioned on surface of membrane do not interact hydrophobic core
    o Found on cytoplasmic side of membrane and form part of cytoskeleton
    o Mad of of mixture of polar and non-polar amino acids
26
Q
  • Hydrophobic nature of membrane = restrictions

- O2 and CO2 necessary and diffuse quickly= good due to their vital role

A
  • Hydrophobic nature of membrane = restrictions

- O2 and CO2 necessary and diffuse quickly= good due to their vital role

27
Q
  • Passive transport:
    o Movement of molecules across membrane without expending chemical energy
    o Molecules move by diffusion
    o Primary mechanism of solute movement
    o Driving force of diffusion = entropy
    o Rate of passive transport depends on concentration gradient
    o Equilibrium = maximum entropy= no net concentration
    o 2 types:
     Simple diffusion
    • Movement of molecules directly across a membrane without involvement of a transporter
    • Rate of simple diffusion depends on molecular size lipid solubility
    • Easily cross membrane: uncharged ions, small molecules, o2 c2 etc.
     Facilitated diffusion
    • Depends on membrane proteins
    • Follows concentration gradients
    • Is specific for certain substances
    • Becomes saturated at high concentrations of transported substance
    • Movement of molecules across membrane with aid of transporter
    • Amino acids, sugars ions and other polar and charged molecules need assistance
    • 2 groups of transport proteins carry out facilitated diffusion (form hydrophilic pathways in membrane)
    o Channel proteins
    o Carrier proteins
    • Aquaporins:
    o Water-specific transport proteins for diffusion of water
    o Narrow channel
    • Gated channel:
    o Transporters can switch between open, closed, intermediate
    o Can be opened or closed based in changes of voltage across membrane or via binding signal molecules
    • Carrier proteins:
    o Each c protein binds a single specific solute and transports it across lipid bilayer (uniport transport)
    o During transport, protein undergoes changes (main difference between carrier and channel)
A
  • Passive transport:
    o Movement of molecules across membrane without expending chemical energy
    o Molecules move by diffusion
    o Primary mechanism of solute movement
    o Driving force of diffusion = entropy
    o Rate of passive transport depends on concentration gradient
    o Equilibrium = maximum entropy= no net concentration
    o 2 types:
     Simple diffusion
    • Movement of molecules directly across a membrane without involvement of a transporter
    • Rate of simple diffusion depends on molecular size lipid solubility
    • Easily cross membrane: uncharged ions, small molecules, o2 c2 etc.
     Facilitated diffusion
    • Depends on membrane proteins
    • Follows concentration gradients
    • Is specific for certain substances
    • Becomes saturated at high concentrations of transported substance
    • Movement of molecules across membrane with aid of transporter
    • Amino acids, sugars ions and other polar and charged molecules need assistance
    • 2 groups of transport proteins carry out facilitated diffusion (form hydrophilic pathways in membrane)
    o Channel proteins
    o Carrier proteins
    • Aquaporins:
    o Water-specific transport proteins for diffusion of water
    o Narrow channel
    • Gated channel:
    o Transporters can switch between open, closed, intermediate
    o Can be opened or closed based in changes of voltage across membrane or via binding signal molecules
    • Carrier proteins:
    o Each c protein binds a single specific solute and transports it across lipid bilayer (uniport transport)
    o During transport, protein undergoes changes (main difference between carrier and channel)
28
Q

o Osmosis: Diffusion of water molecules
 Across a selectively permeable membrane
 From a solution of lower solute concentration to a solution of higher solute concentration
 Selectively permeable membrane must allow water molecules to pass but not solute molecules
 Osmosis can occur in bilayer or from aquaporins
• Hypotonic:
o Higher solute concentration inside of cell than outside
o Water enters via osmosis, cell swells
o Turgor pressure in plant cells due to pressure against cell wall
• Hypertonic:
o Higher solute concentration outside of cell than inside
o Water diffuses out of cell
o Animal and plant cells shrink
• Isotonic:
o Concentration of water inside and outside of cell is equal
o Energy cost: pumping ions form one side to another

A

o Osmosis: Diffusion of water molecules
 Across a selectively permeable membrane
 From a solution of lower solute concentration to a solution of higher solute concentration
 Selectively permeable membrane must allow water molecules to pass but not solute molecules
 Osmosis can occur in bilayer or from aquaporins
• Hypotonic:
o Higher solute concentration inside of cell than outside
o Water enters via osmosis, cell swells
o Turgor pressure in plant cells due to pressure against cell wall
• Hypertonic:
o Higher solute concentration outside of cell than inside
o Water diffuses out of cell
o Animal and plant cells shrink
• Isotonic:
o Concentration of water inside and outside of cell is equal
o Energy cost: pumping ions form one side to another

29
Q
  • Active transport requires energy
    o Form ATP
    o 3 main functions of active transport in cell”
     Uptake of essential nutrients from fluid surrounding cells, even when their concentrations are lower than in cells
     Removal of secretory waste materials from cells or organelles even when concentration of those materials is higher outside the cells or organelles
     Maintenance of essentially constant intracellular concentrations of H+ etc.
    • Charged ions contributes to voltage = creates potential difference aka membrane potential
    o Primary Active Transport:
     The same protein that transports the molecules hydrolyzes ATP to power transport
     Moves positively charged ions across membranes
     H+ pumps (proton pumps)
     Ca2+ pump
     Na+ /K+ pump
    o Secondary Active transport:
     Transport directly driven by ATP
     Transport proteins do not break down ATP
     Instead use a favourable concentration gradient of ions as their energy source
    • Symport
    o Cotransported solute moves through membrane channel in same direction as driving ion
    • Antiport
    o Driving ion moves through membrane channel in one direction, providing energy for active transport of another molecule in opposite direction
A
  • Active transport requires energy
    o Form ATP
    o 3 main functions of active transport in cell”
     Uptake of essential nutrients from fluid surrounding cells, even when their concentrations are lower than in cells
     Removal of secretory waste materials from cells or organelles even when concentration of those materials is higher outside the cells or organelles
     Maintenance of essentially constant intracellular concentrations of H+ etc.
    • Charged ions contributes to voltage = creates potential difference aka membrane potential
    o Primary Active Transport:
     The same protein that transports the molecules hydrolyzes ATP to power transport
     Moves positively charged ions across membranes
     H+ pumps (proton pumps)
     Ca2+ pump
     Na+ /K+ pump
    o Secondary Active transport:
     Transport directly driven by ATP
     Transport proteins do not break down ATP
     Instead use a favourable concentration gradient of ions as their energy source
    • Symport
    o Cotransported solute moves through membrane channel in same direction as driving ion
    • Antiport
    o Driving ion moves through membrane channel in one direction, providing energy for active transport of another molecule in opposite direction
30
Q
  1. 6 Exocytosis and endocytosis
    - Eukaryotic cells import larger molecules via endo and exocytosis
    - Both processes require ATP
A
  1. 6 Exocytosis and endocytosis
    - Eukaryotic cells import larger molecules via endo and exocytosis
    - Both processes require ATP
31
Q

Exocytosis:
- Releases molecules to the outside via secretory vesicles
o Secretory vesicles move through cytoplasm and contact plasma membrane + release contents to cell exterior
 Animal: cells secrete glandular and peptide hormones
 Plants: secrete carbohydrates to build strong cell wall

A

Exocytosis:
- Releases molecules to the outside via secretory vesicles
o Secretory vesicles move through cytoplasm and contact plasma membrane + release contents to cell exterior
 Animal: cells secrete glandular and peptide hormones
 Plants: secrete carbohydrates to build strong cell wall

32
Q

Endocytosis:
- Brings materials into cells via endocytic vesicle
- Two phases:
o Bulk phase endocytosis:
 extracellular water is taken in among any molecules happening to be in solution with the water (0 binding of surface receptors)
o Receptor-mediated endocytosis:
 molecules taken in are bound to outer cell surface via receptor proteins (integral proteins of plasma membrane), bind certain molecules, create coated pit (formed form network proteins called clathrin – reinforce cytoplasmic side)
 pit pinches free form plasma membrane – forms endocytic vesicle and loses clathrin coat and fuses with lysosome
 enzymes in lysosome digests contents of vesicle, breaks down molecules into small bits
 Small molecules enter vesicle membrane via transport proteins
 Phagocytosis : form of endocytosis
Two processes cycle membrane segments between internal cytoplasm and cell surface = maintains surface area of plasma membrane @ controlled levels

A

Endocytosis:
- Brings materials into cells via endocytic vesicle
- Two phases:
o Bulk phase endocytosis:
 extracellular water is taken in among any molecules happening to be in solution with the water (0 binding of surface receptors)
o Receptor-mediated endocytosis:
 molecules taken in are bound to outer cell surface via receptor proteins (integral proteins of plasma membrane), bind certain molecules, create coated pit (formed form network proteins called clathrin – reinforce cytoplasmic side)
 pit pinches free form plasma membrane – forms endocytic vesicle and loses clathrin coat and fuses with lysosome
 enzymes in lysosome digests contents of vesicle, breaks down molecules into small bits
 Small molecules enter vesicle membrane via transport proteins
 Phagocytosis : form of endocytosis
Two processes cycle membrane segments between internal cytoplasm and cell surface = maintains surface area of plasma membrane @ controlled levels

33
Q

Pathways:

  1. Reception:
    a. Binding of signal molecule with specific receptor on target cells
    b. Most receptors found on plasma membrane, can also be found on internal membranes (ER)
    c. Other receptors: soluble proteins found in cytoplasm
A

Pathways:

  1. Reception:
    a. Binding of signal molecule with specific receptor on target cells
    b. Most receptors found on plasma membrane, can also be found on internal membranes (ER)
    c. Other receptors: soluble proteins found in cytoplasm
34
Q
  1. Transduction:
    a. Process where signal reception triggers changes within cell to cause cellular response
    b. Cascade of reactions including several diff. molecules (signalling cascade)
A
  1. Transduction:
    a. Process where signal reception triggers changes within cell to cause cellular response
    b. Cascade of reactions including several diff. molecules (signalling cascade)
35
Q
  1. Response:
    a. Transduction signals causes specific cellular response
    b. Diff. signalling pathways = different downstream responses (eg: activation of specific enzyme causes changes in gene expression)
A
  1. Response:
    a. Transduction signals causes specific cellular response
    b. Diff. signalling pathways = different downstream responses (eg: activation of specific enzyme causes changes in gene expression)
36
Q

Signal reception triggers response pathways in cell (Transduction)
- Binding activates cascade
- Common characteristic of signalling mechanisms: signal is relayed inside cell via protein kinases (enzymes transferring phosphate group from ATP to sites of proteins)
- Protein Kinases: only active when surface receptor binds to a signal molecule
- Protein Kinases acts in chain called phosphorylation cascade
o Last protein in cascade is known as target protein
o This protein is stimulated or inhibited
o Protein phosphatases: remove phosphate groups from target proteins, always active in cells
- Characteristic of signal transduction pathways: amplification:
o Increase in magnitude of each step as transduction pathway proceeds
o Occurs when proteins carrying out pathway are enzymes
o More enzymes-catalyzed steps in reaction = greater amplification = few extracellular signal molecules binding to receptors can proceed full internal response

A

Signal reception triggers response pathways in cell (Transduction)
- Binding activates cascade
- Common characteristic of signalling mechanisms: signal is relayed inside cell via protein kinases (enzymes transferring phosphate group from ATP to sites of proteins)
- Protein Kinases: only active when surface receptor binds to a signal molecule
- Protein Kinases acts in chain called phosphorylation cascade
o Last protein in cascade is known as target protein
o This protein is stimulated or inhibited
o Protein phosphatases: remove phosphate groups from target proteins, always active in cells
- Characteristic of signal transduction pathways: amplification:
o Increase in magnitude of each step as transduction pathway proceeds
o Occurs when proteins carrying out pathway are enzymes
o More enzymes-catalyzed steps in reaction = greater amplification = few extracellular signal molecules binding to receptors can proceed full internal response

37
Q

sterols function

A

used to regulate

38
Q

are alpha helix channels hydrophilic or hydrophobic

are they efficient

A

hydrophobic

yes

39
Q

advantage of enzymatic proteins embedded side-by-side in plasma membrane?

A

facilitates several steps in same enzymatic pathways being spatially linked

40
Q

Passive transport is based on diffusion what does that mean for concentration gradients

A

goes from high to low

41
Q

K+ Voltage Gated Channel

A

every single ion possesses a voltage gated channel specific for it
important for fast muscle contraction must build up high concentration gradients
very selective

42
Q

carrier proteins

A

amoutn differes based on different metabolicc rates

43
Q

why would a plant in normal conditions create a greater solute concentrations within plants relative to the extracellular solution

A

promote water uptake, maintains tension (turgor pressure), increase cellular streaming for structural support

44
Q

difference energy wise between carrier and active transport

A

carrier = no atp

active needs atp

45
Q

how are H+ pumps, Ca2+, Na and K pump able to create membrane potential

A

by creating concentration gradients

46
Q

Active transport of ions creates what

A

voltage

47
Q

What is membrane potential

A

electricxal potential diffference across plasma membrane

48
Q

The Na+ and K+ pump

A

creates e;ectrochrmical gradient, stored energy used for other transport mechanisms, uself in many physiological processes

49
Q

Symport

A

ATP needed, with concentration gradient

50
Q

Antiport

A

Against concentration gradient

51
Q

is bulk phase selective, are cathrins involved

A

no, and no

52
Q

clathrins

A

protein layer, adds extra support to plasma membrane, receives signal for protein receptors and collapses into the cell, initiates collapse and creation of vesicle

53
Q

phagocytosis is a form of

A

endocytosis

54
Q

Role of membrane in cell singalling

A

sensing and responding to changes in the environment

55
Q

phosphorylation cascade

A

domino, cascade effect, protein phosphate remove phosphate group of switch signals, phosphorylation cascades phosphorylate to phosphorylate