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M2M Unit III > Week 3 stuff > Flashcards

Week 3 stuff Flashcards

1
Q

Regulation of Microtubules

A
  1. Microtubule capping proteins

2. Microtubule severing proteins

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2
Q

3 examples of microtubule severing proteins

A

katanin
spastin
fidgetin

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3
Q

Microtubule functions (4)

A
  1. Cellular cytoskeleton
  2. intracellular transport
  3. cell division
  4. cilia
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4
Q

Microtubules in cell division (3)

A

astral microtubules
kinetochore microtubules
centrospindle- overlapping attachment

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5
Q

Domains of microtubules

A

alpha, beta, and gamma

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6
Q

Plus end Subunit

A

beta

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7
Q

minus end subunit

A

alpha

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8
Q

GTP cap function

A

Stabilization

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9
Q

Microtubule organizing center

A

centrosome

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10
Q

Each centrosome contains 2 ___

A

centrioles

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11
Q

Gamma tubulin forms _____ which binds to ____

A

rings outside of centriole

binds to minus ends

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12
Q

Kinesin homodimer

A

Kinesin I

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13
Q

Kinesin heterodimer

A

Kinesin II

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14
Q

N terminus of kinesin

A

head domain

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15
Q

C terminus of kinesin

A

tail domain

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16
Q

for the tail domain to bind to cargo there is usually an associated ____

A

adaptor protein

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17
Q

Energy used in kinesins

A

ATP

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18
Q

Retrograde transport of ___ is essential for neuron proliferation

A

NGF

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19
Q

In neuropathies, damage to the forward pathway is typically ____ than damages to the retrograde pathway

A

less severe

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20
Q

Actin energy

A

ATP

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21
Q

Minus end of actin has ____

A

open ATP pocket

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22
Q

Actin step needed to grow

A

nucleation

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23
Q

Actin filament structure and regulation (4)

A
  1. G actin concentration
  2. ADP to ATP exchange
  3. Capping
  4. Depolymerization/severing
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24
Q

2 proteins that regulate nucleation

A

Formin (FH2)

Arp2/3

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25
Q

Arp2/3

A

branched

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26
Q

FH2

A

bundles

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27
Q

Arp2/3 and FH2 initiate nucleation by forming _____

A

pseudonucleation centers

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28
Q

Formin activation

A

small GTPases bind and activate

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29
Q

Actin roles in cell function

A
  1. epithelial cell polarity
  2. contraction
  3. cell motility
  4. cell division
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30
Q

Tight Junctions (TJ)

A

Separate apical and basolateral

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31
Q

Microvilli-> ___ dependent

A

FH2

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32
Q

Myosin involved in transport

A

Unconventional myosins

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33
Q

Cell motility: Arp2/3 complex regulates ____ formaion

A

lamellipodia

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34
Q

______ complex regulates lemellipodia formation

A

Arp2/3

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35
Q

Formins complex regulate ____ formation

A

filopodia

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36
Q

_____ regulate filopodia formation

A

Formins complex

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37
Q

Function of intermediate filaments

A

mechanical stability

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38
Q

receptor lipophilic

A

can be intracellular

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39
Q

receptor hydrophilic

A

has to be on cell surface

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40
Q

cell storage- lipophilic

A

NO

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41
Q

cell storage- hydrophilic

A

yes in vesicles

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42
Q

control of release lipophilic

A

only by synthesis

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43
Q

speed lipophilic

A

slow

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44
Q

speed hydrophilic

A

fast

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45
Q

Receptor types (4)

A
  1. Ligand (or voltage) gated ion channels
  2. GPCRs
  3. Enzyme linked receptors
  4. Nucleat receptors- TFs activated by cell penetrating signaling molecules
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46
Q

Second messenger examples (5)

A
  1. Ca2+
  2. cAMP
  3. IP3
  4. DAG
  5. NO
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47
Q

cAMP is generated by ___

A

Adenylate cyclase (AC)

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48
Q

DAG are generated by ___

A

Phospholipase C (PLC)

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49
Q

NO generated by ____

A

NOS

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50
Q

Signaling proteins, activated by receptors or second messengers, can activate other signaling or effector proteins (including enzymes that generate second messengers) in a variety of ways. Including:

A

protein modification

protein-protein binding

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51
Q

Amplification often depends on _____

A

how many downstream targets affected

time it takes to terminate activity of a signaling molecule

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52
Q

signaling- Node

A

multiple input/multiple outputs

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53
Q

signaling- module

A

groups that function together

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54
Q

PDE5 function

A

breaks down cyclic GMP to GMP

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55
Q

PDE5 binding

A

catalytic site and noncatalytic site

cooperative binding

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56
Q

Protein that phosphorylates PDE5

A

PKG

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57
Q

PDE5 inhibitor

A

Sildenafil (Viagra)

binds to catalytic site

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58
Q

Receptor tyrosine kinase activation is driven by _____

A

ligand mediated dimerization

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59
Q

Key downstream activation of EGFR

A

Ras

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60
Q

Active Ras

A

Ras-GTP

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61
Q

Inactive Ras

A

Ras GDP

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62
Q

GTP->GDP

A

GAP

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63
Q

GDP->GTP

A

GEF

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64
Q

Adaptor protein involved in Ras activation

A

Grb2

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65
Q

Domains of Grb2

A

SH2 and SH3 domains

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66
Q

SH2 domain of Grb2 binds____

A

Phospho-Tyr containing peptides

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67
Q

SH3 domain of Grb2 binds ____

A

Pro-containing peptides (Sos)

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68
Q

Ras GEF involved in Ras activation

A

Sos

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69
Q

Recruitment of Sos to the receptor ____

A

brings Sos in close proximity with Ras which is sequestered at the cell membrane

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70
Q

Two ways to inhibit EGFR

A
  1. small molecule inhibitors that bind kinase domain

2. antibodies that bind extracellular domain

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71
Q

Example of antibody that blocks ligand binding site

A

cetuximab

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72
Q

Example of small molecule inhibitor of EGFR

A

getfitinib

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73
Q

What part of EGFR does getfitinib inhibit

A

ATP binding domain

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74
Q

Mutant EGFR is more active and more potently inhibited by ____

A

getfitinib

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75
Q

______ mutations in EGFR cause resistance to getfitinib

76
Q

G-protein coupled receptors structure

A

7 helical transmembrane domains

Inside of cell:
i2 loop (bt 3 and 4)
i3 loop (5 and 6)
C-tail

77
Q

____ % of all non antibiotic prescription drugs act through G protein coupled receptors

78
Q

G proteins that interact with GPCRs

A

Galpha

Beta-Gamma

79
Q

G alpha is inactive when bound to ___

80
Q

G alpha is inactivated when bound to ___

81
Q

Effector regulation following GPCR activation is mediated by ____

A

free alpha- GTP and beta-gamma subunits

82
Q

bordello pertussis effect on GPCR

A

ribosylates Galpha near C terminal and locks it in the inactive state

83
Q

bacteria that inactivates GPCR

A

bordello pertussis (whooping cough)

84
Q

bacteria that activates GPCR

85
Q

cholera effect on GPCR

A

ribosylates Galpha near GTP binding site to inhibit GTPase activity

86
Q

Adrenergic receptors

A

Beta AR

Alpha AR

87
Q

Muscarinic receptors

A

m2 AchR and m3AchR

88
Q

BetaAR and m2AchR- effects

A

cAMP production
BetaAR- stimulates
m2AchR- inhibits

89
Q

alphaAR and m3 AchR- efects

A

stimulate Phospholipase C, Ca, and lipid signals

90
Q

Agonists of beta adrenergic receptor

A

norepinephrine, epinephrine, or isoproterenol

91
Q

Antagonists of beta adrenergic receptor

A

propranolol or metoprolol

92
Q

cAMP->____->______

A

PKA->ca channels

93
Q

Increase of calcium in hear causes_____

A

increased heart rate and contraction

94
Q

Agonists of alpha adrenergic receptor

A

norepinephrine, epinephrine, or phenylephrine

95
Q

Antagonists of alpha adrenergic receptor

96
Q

Agonist of m2 muscarinic cholinergic receptors

A

acetylcholine, Muscarine

97
Q

Enzymes that catalyze cAMP to AMP

A

Phosphodiesterases

98
Q

General PDE inhibitors

A

caffeine

theophyline

99
Q

cAMP specific PDE inhibitors

A

Milinirone PDE3

Rolipram PDE4

100
Q

Proteins associated with GPCR desensitizaiton

A

GRK

Beta arrestin

101
Q

After beta arrestin binds to GPCR ____

A

The receptor is internalized (endocytosed) and targeted for degradation in the lysosome

102
Q

Beta arrestin b-arrestin also couples GPCRs to activation of additional downstream signaling pathways such as the ____ and ___ and ____ pathways

A

JNK
ERK
MAP kinase

103
Q

More kinases or phosphatases?

104
Q

a kinase catalyzes what reaction?

A

nucleophilic attack of hydroxyl group onto gamma phosphate of an ATP molecule

105
Q

Kinase domain

where does atp bind?

A

small and large lobe

cleft between lobes

106
Q

Lobe of kinase that interacts with substrate

107
Q

closed conformation of kinase

A

glycine rich loop in the small lobe forces the gammaPhosphate of the ATP into the right position for phosphorylation (a fast reaction

108
Q

Open conformation of kinase

A

glycine rich loop then allow exchange of the generated ADP for a new ATP (a slow reaction).

109
Q

Which conformation of kinase is conserved? Why is this a problem?

A

open

specific inhibitors

110
Q

Important immunosupressants

A

calcineurin and mTOR inhibitors

111
Q

Rate limiting step of GPCR activation

A

GDP dissociation

112
Q

Beta adrenergic receptor G protein

113
Q

Alpha adrenergic receptor G protein

114
Q

m2 muscarinic cholergenic receptor G protein

115
Q

m3 muscarinic receptor G protein

116
Q

Desensitization of GPCRs (4 steps)

A
  1. GRK interacts w/ beta gamma and phosphorylates receptor
  2. Beta arrestin binds to phosphorylated receptor and inhibits alpha from rebinding
  3. internalization
  4. degradation by lysosome
117
Q

Calcium sources (4)

A
  1. ECF
  2. ER/SR
  3. Mitochondria
  4. Nuclear envelope
118
Q

Ca Ion channels- plasma membrane (2)

A
  1. Voltage and ligand gated Ca channels

2. Store operated Ca channels (Orai1)

119
Q

Ca ion channels ER/SR (2)

A
  1. IP3 receptors
  2. Ryanodine receptors
    Ca moves from lumen to cytoplasm
120
Q

Movement of Ca in mitochondria

A

depends on Ca gradient

can be sink or source

121
Q

Two types of Ca transporters

A
  1. Ca pumps (ATP)

2. Na/Ca exchangers

122
Q

Two types of Ca pumps

A
  1. PMCA- Cyt-> extracellular space

2. SERCA- into lumen of ER

123
Q

NCX exchange

124
Q

Cytoplasmic buffer

A

parvalbumin

125
Q

Cytoplasmix buffers restrict the ____ and ____ spread of calcium and also serve as ____

A

spatial
temporal
temporary storage sites for calcium

126
Q

Example of ER ca buffer

____ capacity _____ affinity

A

calsequestrin

high, low

127
Q

Ca effectors

A
  1. membrane potential
  2. PKC
  3. Synaptotagmin
  4. Calmodulin
128
Q

function of synaptotagmin

A

Ca dependent fusion of synaptic vesicles

129
Q

Binding of Ca causes ___ domains of ___ and _____ to associate with the plasma membrane

A

C2 domains
PKC
Synaptotagmin

130
Q

Structure of calmodulin

A

4 EF-hand binding sites

Each ca coordinated with 5 oxygens

131
Q

Steps of helper T lymphocyte activation

A
  1. antigen binds to TCR
  2. Activate tyrosine kinase
  3. Activate phospholipase C
  4. PIP2-> IP3 and DAG
  5. . Release ca, depleting ER stores
  6. STIM1 senses depleted stores-> Orai1
  7. Ca-calmodulin binds to calcineurin
  8. Dephosphorylates NFAT
  9. NFAT translocates to nucleus and promotes expression of IL-2 gene
132
Q

True intestinal stem cell

A

Crypt base columnar cells (CBC)

133
Q

Important gene expressed in CBCs

134
Q

Niche for Lgr5 stem cells

A

Paneth cells

135
Q

Epidermolysis bullosa is a defect in ____

A

Collagen 7

136
Q

______ is a defect in collagen 7

A

Epidermolysis bullosa

137
Q

Needed for conversion of somatic cells to ES-like stem cells

A

Reprogramming factors

138
Q

4 reprogramming factors

A
  1. Oct3/4
  2. Sox2
  3. c-Myc
  4. Klf4
139
Q

New therapies in cancer treatment

A

Directed against cancer stem cells

140
Q

PSA in prostate cancer

A

Prostate specific antigen

lower levels are better

141
Q

Methods of hormone therapy in prostate cancer

A
  1. Surgical bilateral orchiectomy
  2. GNRH agonis
  3. Anti- androgen
142
Q

Hormone refractory prostate cancer

A

Disease progression with a testosterone of less than 50 ng.dl

143
Q

For most mammalian cell types, adhesion to ______ or the ____ is critical for cell survival

A

other cells

Extracellular matric

144
Q

Cell adhesion is ___

145
Q

The relative amount of ECM in different tissues ____ , from connective tissues, where the ECM may occupy_____ to brain, where it is ________.

A

Varies greatly
the bulk of the volume
a very minor component only

146
Q

Functions of the ECM

A
  1. scaffold for cells

2. regulation of various cell functions

147
Q

In connective tissues, the ECM macromolecules are synthesized and secreted primarily by______ or ______, such as ______ (in cartilage) or _______ (in bone).

A

Fibroblasts
specialized members of the fibroblast family
chondroblasts
osteoblasts

148
Q

4 major classes of molecules in ECM

A
  • glucosaminoglycans (GAGs), linked to proteins called proteo glycans
  • fibrous proteins (collagen, elastin)
  • multidomain adaptor proteins (fibronectin, Laminin)
  • water and many solutes
149
Q

Proteoglycans form ___

A

highly hydrated gel in which fibrous and multidomain proteins are embedded

150
Q

Fibrous proteins give ECM ___

A

mechanical properties

151
Q

Structure of GAGs

A

unbranched polysachharide composed of disaccharide repeats

  • one is always amino sugar, usually sulfated
  • the other is typically uronic acid
152
Q

GAGs charge

A

negative

Gives capacity to be highly hydrated

153
Q

The “core protein” of proteoglycans has attached to ___ special ______, and these serve as the _____ for polysaccharide assembly

A

serines
link tetrasaccharides
primers

154
Q

Some PGs are membrane-bound molecules, anchored via a ______ or linked via a ______

A

transmembrane core protein tail

GPI anchor

155
Q

Collagen I

A

most common form

abundant in connective tissues

156
Q

Collagen IV

A

characteristic of basal lamina

157
Q

Elastin

A

Provides elasticity

158
Q

Multidomain adaptor protein

A

act as binding sites for other matrix macromolecules and for adhesion molecules on the surfaces of cells.

159
Q

Fibronectin structure

A

large, dimeric glycoprotein

large subunits linked together by disulfide bonds

160
Q

Type III fibronectin repeat domain binds to ___

161
Q

Binding sequence of Type III fibronectin repeats

A

Arg-Gly-Asp

162
Q

secreted fibronectin assembles in the ECM into highly insoluble _____

A

fibronectin fibrils

163
Q

Three subunits of laminin

A

alpha, beta, gamma

164
Q

Lamina is found in ___

A

basal lamina only

165
Q

Laminins, like fibronectins, have numerous binding sites for cells (integrins; see below) and other _____ that link them to ____

A

ECM proteins

Collagen IV

166
Q

Proteins that regulate the ECM

167
Q

Proteases in ECM

A 
matrix metalloproteases (MMP)
can be selective or promiscuous
168
Q

Extracellular protease activity is particularly important in _____ and in __________

A

tissue remodelind in development

cell invasion of tissues

169
Q

When bound to a ligand, CAMs signal their engagement via _____, thus affecting _____

A

conformational change to the cell interior

cell function

170
Q

3 most abundant classes of CAMS

A
  1. Cadherins
  2. IgSF CAMS
  3. Integrins
171
Q

Cadherin binding is ___ dependent

172
Q

Cadherins bind to other cells via cadherins on the juxtaposed cell surface (homophilic binding) in a________

A

zipper like fashion

173
Q

Cadherin structure

A

single pass transmembrane glycoproteins that operate as homodimers

174
Q

IgSF members do not form ____, and binding does not require ____.

A

dimers

Ca2+

175
Q

2 homophilic CAMs

1 heterophilic CAM

A

Homo- cadherin, IgSF CAM

hetero- integrins

176
Q

Structure of integrins

A

alpha/beta heterodimers

both participate in ligand binding

177
Q

Typical ligands of integrins

A

laminin
fibronectin
collagen
Many involving RGD sequence

178
Q

Cytoskeletal link of CAMs

A

acin-binding proteins

catenins

179
Q

In addition to providing the cytoskeletal link, CAM-associated proteins also are involved in the ____, _____, and in _____

A

regulation of adhesion
control of actin polymerization
cell signaling

180
Q

Fibronectin null mutant mice are ______

A

early embryonic lethal

181
Q

Some laminin mutations have been linked to _____ (glomerular filtration defect) and to _______ problems in children

A

nephrotic syndrome

neuromuscular junction/muscle innervation problems

182
Q

Loss-of-function mutations of matrix metalloproteases (MMP) 2 or 13 cause

A

inherited osteolysis/arthritis syndromes and bone dysplasias

183
Q

Overexpression of some MMPs, specifically MMP_, MMP_ and MMP__, correlates

A

2,9,14

high invasiveness and poor prognosis in many cancers

184
Q

Two types of leukocyte adhesion deficiencies

A

Type I- integrin

Type II- selectin

185
Q

One of the early signs of carcinogenesis is a change in CAM(s). This is accompanied by
(and may be the reason for) perturbed_________

A

cell polarity and cytoarchitecture

186
Q

Cadherin down-regulation appears to be a prerequisite for the dispersal of ________

A

endothelium derived cancer cells

M2M Unit III (3 decks)

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