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Biochem > cellular > Flashcards

cellular Flashcards

1
Q

regulators of cell cycle

A

cyclins, CDKs, tumor suppressors

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2
Q

shortest phase of cell cycle

A 
MITOSIS:
prophase.
metaphase.
anaphase.
telophase.
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3
Q

cyclins

A

regulatory proteins.
phase-specific.
ACTIVATE CDKs.

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4
Q

CDKs (cyclin-dependent kinases)

A

constitutive and inactive.

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5
Q

tumor suppressors

A

Rb and p53.
inhibit G1 to S progression.

*mutations = unrestrained growth

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6
Q

permanent cells

A

remain in G0.
regenerate from STEM CELLS.

neurons.
skeletal m.
cardiac.
RBCs.

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7
Q

stable (quiescent) cells

A

enter G1 from G0 when stimulated.

hepatocytes.
lymphocytes.

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8
Q

labile cells

A

never go to G0.
rapidly divide with short G1.

BM.
gut epith.
skin.
hair follicles.

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9
Q

RER

A

site of synth of secretory (EXPORTED) proteins.

N-linked oligosaccharide addition to many proteins.

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10
Q

which cells are rich in RER?

A

mucus-secreting GOBLET CELLS of small intestine.

Ab-secreting PLASMA CELLS.

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11
Q

Nissl bodies

A

RER of neurons-

synthesize enzs and peptide NTs.

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12
Q

free ribosomes

A

unattached to any memb.

synth of cytosolic and organellar prots.

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13
Q

SER

A
  1. steroid synth (hormones).
  2. detox of drugs, poisons.
  3. carb metabolism.
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14
Q

which cells are rich in SER?

A

liver hepatocytes.

steroid-producing cells of adrenal cortex.

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15
Q

peroxisome

A

catabolism of VLCFA (very long chain fatty acids) and amino acids

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16
Q

proteasome

A

barrel-shaped protein complex.

degrades damaged/unnecessary proteins tagged with UBIQUITIN for destruction.

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17
Q

Golgi apparatus

A

distribution center for proteins and lipids-

from ER to plasma memb and vesicles.

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18
Q

how does Golgi change asparagine?

A

modifies N-oligosaccharides

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19
Q

how does Golgi change serine and threonine?

A

adds O-oligosaccharides

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20
Q

mannose 6-phosphate

A

Golgi adds mannose 6-phosphate to proteins for trafficking to LYSOSOMES

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21
Q

endosome

A

sorting center for material from outside cell or from Golgi - sends it to lysosomes for destruction or back to memb/Golgi for further use

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22
Q

vesicular trafficking prot: COP I

A

retrograde.

Golgi to ER.

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23
Q

vesicular trafficking prot: COP II

A

anterograde.

RER to cis-Golgi.

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24
Q

vesicular trafficking prot: clathrin

A

trans-Golgi
to
lysosomes, memb
to endosomes (receptor-mediated endocytosis)

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25
Q

I cell disease

A

(inclusion cell disease)
inherited lysosomal storage disorder.

fail to add mannose 6-phosphate tag to lysosome proteins = enzs secreted outside of cell instead of being targeted to lysosomes.

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26
Q

features of I cell disease

A

coarse facial features.
CLOUDED corneas.
restricted joint mvmt.
high plasma levels of lysosomal enzymes.

*often fatal in childhood

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27
Q

microtubule

A

cylindrical structure.

helical array of polymerized dimers (ALPHA and BETA TUBULIN) - each dimer has 2 GTP.

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28
Q

microtubules are incorporated into…?

A

flagella (dynein).
cilia (dynein).
mitotic spindles.
axoplasmic transport in neurons*

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29
Q

molecular motor proteins

A

transport cellular cargo (organelles, intracellular vesicles) toward opposite ends of microtubule tracks.

use ATP.

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30
Q

dynein

A

Retrograde to microtubule (+ to -)

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31
Q

kinesin

A

Anterograde to microtubule (- to +)

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32
Q

drugs that act on microtubules

A 
Mebendazole, thiabendazole.
Colchicine.
Griseofulvin.
Vincristine, vinblastine.
Paclitaxel.

“Microtubules Can Grow Very Progressively”

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33
Q

Chediak-Higashi syndrome

A

microtubule polymerization defect =
decreased fusion of phagosomes and lysosomes.

results in recurrent pyogenic infxs, partial albinism, peripheral neuropathy.

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34
Q

cilia

A

9+2 arrangement of microtubules.

contains dynein.

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35
Q

dynein in cilia

A

axonemal dynein.

ATPase links peripheral 9 doublets and causes bending of cilium by differential sliding of doublets.

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36
Q

Kartagener’s syndrome

A

IMMOTILE CILIA due to dynein arm defect.

results in male and female infertility (sperm immotile), bronchiectasis, recurrent sinusitis.
assoc. with situs inversus.

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37
Q

cytoskeleton: actin, myosin

A

microvilli.
muscle contraction.
cytokinesis.
adherens junctions (intermediate).

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38
Q

cytoskeleton: microtubules

A

movement:

cilia.
flagella.
mitotic spindle.
axonal trafficking.
centrioles.
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39
Q

cytoskeleton: intermediate filaments

A

structure:

vimentin.
desmin.
cytokeratin.
lamins.
GFAP.
neurofilaments.
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40
Q

plasma membrane

A 
asymm lipid bilayer:
cholesterol 50%.
phospholipids 50%.
sphingolipids.
glycolipids.
proteins.
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41
Q

high chol or long saturated FA content of plasma memb means….?

A

increased melting temp.

decreased fluidity.

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42
Q

vimentin

A

conn tissue

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43
Q

desmin

A

muscle

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44
Q

cytokeratin

A

epith cells

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45
Q

GFAP

A

neuroGlial cells (astrocytes)

46
Q

neurofilaments

A

neurons

47
Q

Na-K ATPase

A

ATP site on cytoplasmic membrane:
1 ATP consumed.
3 Na out.
2 K in.

48
Q

ouabain

A

inhibits Na-K ATPase by binding to K site

49
Q

cardiac glycosides (digoxin, digitoxin)

A

directly inhibit Na-K ATPase
= indirect inhibition of Na/Ca exchange
(increase intracellular Ca = increase cardiac contractility)

50
Q

most abundant protein in human body?

A

collagen

51
Q

collagen function

A

organizes and strengthens ECM

52
Q

type I collagen

A 
90%.
bone
skin
tendon
dentin
fascia
cornea
late wound repair (scars)
53
Q

type I collagen defect in…

A

osteogenesis imperfecta

54
Q

type II collagen

A

cartilage (including hyaline)
vitreous body
nucleus pulposus

55
Q

type III collagen

A 
AKA reticulin.
skin
blood vessels
lungs
intestine
RES
uterus
fetal tissue
granulation tissue
56
Q

type III collagen defect in…

A

Ehlers-Danlos

57
Q

type IV collagen

A

basement memb

or basal lamina

58
Q

type IV collagen defect in…

A

Alport syndrome

59
Q

collagen formation step 1: synthesis

A

in RER:
translate collagen alpha chains (preprocollagen).

usually GLY-X-Y polypeptide where
X and Y are proline and lysine.

60
Q

collagen formation step 2: hydroxylation

A

in RER:
hydroxylate specific proline and lysine residues.

REQUIRES VIT C

61
Q

collagen formation step 3: glycosylation

A

in RER:
glycosylate pro-alpha-chain hydroxylysine residues.

form PROCOLLAGEN via hydrogen and disulfide bonds (TRIPLE HELIX of 3 pro-alpha-chains)

62
Q

collagen formation step 4: exocytosis

A

procollagen triple helix exocytosed into extracellular space

63
Q

collagen formation step 5: proteolytic processing

A

outside fibroblast:

cleave terminal regions of procollagen to transform into insoluble TROPOCOLLAGEN

64
Q

collagen formation step 6: cross-linking

A

outside fibroblast:
reinforce numerous tropocollagen molecules with covalent lysine-hydroxylysine cross-linkage by LYSYL OXIDASE to make COLLAGEN FIBRILS

65
Q

what step in collagen formation does scurvy affect?

A

vit C deficiency — affects hydroxylation of proline and lysine residues

66
Q

osteogenesis imperfecta

A

genetic bone disorder (BRITTLE BONE) caused by various gene defects.

most commonly auto dom with abnormal collagen type I.

67
Q

features of osteogenesis imperfecta

A
  1. multiple fractures with minimal trauma.
  2. BLUE SCLERA due to translucency of conn tissue over choroid.
  3. hearing loss due to abn middle ear bones.
  4. dental imperfections due to lack of dentin.

may be confused with child abuse.

68
Q

which form of OI is most fatal?

A

type II - in utero or neonatal period

69
Q

Ehler-Danlos syndrome

A

faulty collagen synthesis.

varying inheritance and severity.
auto dom or recessive.

70
Q

what type of collagen is most often affected in Ehler-Danlos syndrome?

A

type V (classic syndrome)

*6 types total

71
Q

features of Ehler-Danlos syndrome

A
  1. hyperextensible skin.
  2. tendency to bleed (easy bruising).
  3. hypermobile joints.

may be assoc with joint dislocation, berry aneurysms, organ rupture.

72
Q

Alport syndrome

A

due to various gene defects leading to abnormal TYPE IV collagen.

most common is X-linked recessive.

73
Q

features of Alport

A
  1. progressive hereditary nephritis
  2. deafness
  3. ocular disturbances

*type IV important struct component of BM in kidney, ears, eyes

74
Q

elastin

A

stretchy protein in lungs, large arteries, elastic ligaments, vocal cords, ligamenta flava.

75
Q

ligamenta flava

A

connect vertebrae: relaxed and stretched conformations

76
Q

elastin is rich in which AA?

A 
proline.
glycine.
lysine.
alanine.
valine.

(nonpolar, non-hydroxylated forms)

77
Q

tropoelastin

A

elastin precursor.

has fibrillin scaffolding.

78
Q

what breaks down elastin?

A

elastase

79
Q

what inhibits elastase?

A

alpha 1 antitrypsin

80
Q

wrinkles of aging

A

due to reduced collagen and elastin prod.

81
Q

Marfan’s syndrome

A

defect in fibrillin

82
Q

emphysema

A

can be caused by alpha 1 antitrypsin deficiency resulting in EXCESS ELASTASE activity

83
Q

PCR purpose

A

amplify desired fragment of DNA

84
Q

PCR steps

A
  1. denaturation.
  2. annealing.
  3. elongation.

*repeated multiple times for DNA sequence amplification (25-35 cycles)

85
Q

PCR: denaturation

A

DNA denatured by heating to generate 2 separate strands

86
Q

PCR: annealing

A

during COOLING, excess premade DNA primers anneal to specific seq on each strand to be amplified

87
Q

PCR: elongation

A

heat-stable DNA polymerase replicates the DNA seq that follows each primer

88
Q

components needed for PCR

A
  1. DNA template (seq may be unknown).
  2. two primers complementary to regions flanking DNA template.
  3. heat-stable DNA polymerase that replicates target sequence between primers.
  4. deoxynucleiotide triphosphates to build new DNA.
89
Q

agarose gel electrophoresis after PCR

A

for SIZE separation of PCR products (small molecules travel further).

compared against DNA ladder.

90
Q

Southern blot

A

DNA

91
Q

Northern blot

A

RNA

92
Q

Western blot

A

protein

93
Q

Southwestern blot

A

DNA-binding proteins (TFs, nucleases, histones)

94
Q

microarray

A

thousands of nucleic acid sequences arranged in grids on glass or silicon.
DNA/RNA probes are hybridized to chip.
scanner detects relative amts of COMPLEMENTARY BINDING.

95
Q

purpose of microarray

A

to profile gene expression levels of thousands of genes simultaneously.

can detect SNPs.

96
Q

ELISA

A

enzyme-linked immunosorbent assay.

rapid immunologic test for AG-AB REACTIVITY.

97
Q

ELISA with test Ag

A

Ag coupled to color-generating enzyme.
see if imm system recognizes Ag.

(if target is acquired in sample, test soln will have intense color rxn to indicate positive result)

98
Q

ELISA with test Ab

A

Ab coupled to color-generating enzyme.
see if a certain Ag is present.

(if target is acquired in sample, test soln will have intense color rxn to indicate positive result)

99
Q

Sn and Sp of ELISA

A

both approach 100% though FP and FN possible

100
Q

FISH

A

fluorescence in situ hybridization.

fluorescent DNA or RNA probe binds specific gene site of interest on chromosomes.

101
Q

FISH use

A

for specific localization of genes.

direct visualization of anomalies.

102
Q

no fluorescence in FISH

A

= gene has been deleted

103
Q

what phase are chromosomes used in karyotyping in?

A

METAPHASE

104
Q

karyotype can be performed with samples of…?

A

blood
BM
amn fluid
placental tissue

105
Q

karyotype use

A

diagnose chromosomal imbalances

106
Q

cloning

A

prod of recombinant DNA molec that is self-perpetuating

107
Q

what is the first component of cloning?

A

euk mRNA of interest

108
Q

what enzyme acts on mRNA in cloning?

A

reverse transcriptase produces cDNA from mRNA

109
Q

what is used to produce a cDNA library?

A

insert cDNA into bacterial plasmids containing antibiotic resistance genes - those that survive on Abx medium produce library

110
Q

transgenic strategies

A

to modify gene expression (in mice) by:

  1. random insertion of gene into mouse genome (constitutive).
  2. targeted insertion or deletion of gene through homologous recombination with mouse gene (conditional).
111
Q

Cre-lox system

A

can inducibly manipulate genes at specific development points using Abx-controlled promoter

112
Q

RNAi

A

dsRNA complementary to mRNA sequence of interest - transfected into humans and promotes degradation of target mRNA (thus knocking down gene expression)

Biochem (7 decks)

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