Metabolism 2.5, 2.6, 7.1, 8.1 Flashcards

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1
Q

What is the role on an enzyme?

A
> speeding up the reaction (catalysts)	
	- no change in reaction 
	- new reaction pathway 
> temperature could be increased 
	- biological systems are sensitive
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2
Q

What is active site?

A
  • enzymes are proteins
    • made often
    • 3D structure
    • structure determines function
  • active site - region where the substrate binds
    • enzyme-substrate complex
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3
Q

What are different models of enzymes?

A
> lock and key 
	- exact fit between enzyme and substrate 
	- 1 enzyme = 1 substrate 
	- products have different shape 
		- formed an released
> induced fit model 
	- proteins can change their shape 
	- substrate binds 
		- induces change in enzyme 
		- active site reshaped 
	- 1 enzyme = many substrates (similar types)
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4
Q

How does an enzyme convert substrate to a product?

A
  • breakdown
    • substrates bind to active site
      • specific bonds exposed
        • stretched (increases likelihood of being broken)
  • synthesis
    • substrates in active site are positioned
      • more likely collision
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5
Q

How do substrates bind to active site?

A
  • collision
    • particles are in motion
    • substrates are smaller so they move faster
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6
Q

What are the factors affecting enzyme activity

A

> enzyme and substrate concentration
- the more enzyme, the faster
- substrate concentration reaches saturation - fixed amount of enzymes
temperature
- the higher, the higher kinetic energy
- too high, enzyme denaturation
pH
- for each reaction there is an optimal pH
- different pH lows the reaction (or prohibits)

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7
Q

What is denaturation?

A
> high temperature or high / low pH
	- active site is altered 
	- no catalyst
> homeostasis of cells is in danger
	- environment is not suitable
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8
Q

What is metabolism?

A
> sum of all reactions that occur in organism to maintain life
> metabolic pathways 
	- sequence of small steps 
	- chain of reactions 
	- sometimes cycle
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9
Q

What are types of inhibitors?

A
  • inhibitors reduce enzyme activity
  • competitive inhibitor
    • similar to substrate
    • active site
    • competes
  • non-competitive inhibition
    • different structure
    • binds to enzyme in different way
    • changes shape of enzyme
  • examples (drugs)
    • saquinavir - HIV protease inhibitor
    • nevirapine - HIV reverse transcriptase inhibitor
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10
Q

How is metabolic pathway regulated?

A
  • end-product inhibition
    • end product of the pathway inhibits metabolism
  • example
    • threonine (initial substrate) binds to theorinine deaminase
    • end product → isoleucine
      • as the concentration increases it binds to allosteric site (non-competitive inhibition)
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11
Q

What are immobilised enzymes?

A
  • enzymes attached to other material (glass, alginate gel)
    • enzyme movement restricted
  • advantages
    • separated from products, stopping reaction at ideal time
    • recycling (cost savings)
    • stability of enzyme increases
      • changes in temp or pH
    • higher concentration of enzyme than dissolved enzyme
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12
Q

Why is lactose-free milk produced?

A
  • lactose → glucose + galactose
  • why?
    • for people who are lactose intolerant
  • glucose and galactose are sweeter
    • less sugar in milk products
    • lactose crystallises during ice cream production
    • quicker fermentation (cheese)
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13
Q

Hershey-Chase experiment

A
  • aim: testing whether proteins or DNA is genetic material
  • viruses injected genetic material
    • radioactive S-35 for proteins
    • radioactive P-32 for DNA
  • infecting bacteria
    • blender to separate non-genetic and genetic material
  • measured radioactivity in supernatant (substance outside) or pellet (genetic material)
  • P-32 in pellet
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14
Q

What is a nucleotide?

A
  • sugar
    • pentose (or different)
  • nitrogenous base
    • 4 types
  • phosphate group
  • formed by forming covalent bonds between phosphate group and sugar
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15
Q

What is the structure of DNA?

A
  • double helix
  • 2 strands of nucleotides linked by covalent bonds
    • different directions
  • complementary base pairing
    • A with T
    • C with G
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16
Q

What are the differences between RNA and DNA?

A
  • different sugars
    • DNA has less oxygen
  • 2 polymers of nucleotides in DNA
  • T → U
17
Q

What is semi-conservative replication?

A
  • base sequence on template determines the sequence on new strand
  • 1 strand parent, 1 strand new
18
Q

Meselson and Stahl experiment

A
  • bacteria grown in N-15 (heavy) medium
    • transferred to N-14
  • samples after 0, 20 and 40 minutes
    • 0 min: heavy and light
    • 20 min (first generation): all intermediate
    • second generation: half intermediate, half light
      = semi-conservative replication
19
Q

What is the first stage of DNA replication?

A
  • helicase unwinds double helix
    • breaking H-bonds
    • group of enzymes
    • use ATP
  • forming replication fork
    • leading strand = towards replication fork
    • lagging strand = opposite
20
Q

What is the second step of DNA replication?

A
  • DNA polymerase III
    • elongates nuclear acid
    • attaches new nucleotides to a stand
    • attaches to 3’ end
      - starts building the chain from 5’
  • RNA primers
    • synthesised by primase
    • start point
      • leading has 1
      • lagging has many = Okazaki fragments
  • DNA strands rewind
21
Q

What is the function of DNA polymerase?

A
  • DNA polymerase III
    • replication in E. coli
    • check
      • polymerases copy bases, check it and cut out if wrong
    • adding nucleotide
      • unless H-bonds are formed, nucleotide is discarded
      • nucleotides are linked by covalent bonds
        • adds to 3’ end
  • DNA polymerase I
    • repair
    • exchange RNA primers into DNA
22
Q

What is the role of nucleosomes?

A
  • eukaryotic DNA in form of chromosomes
    • wrapped around histones
  • 8 histones + DNA = nucleosomes
  • chromosomes can coil
    • packing DNA
23
Q

What is the function of DNA primase?

A
  • generates short RNA primer
    • one on leading strand and maybe on lagging strand
    • extension of nucleotide chain
  • RNA removed by DNA polymerase I (or exonuclease)
24
Q

What is the function of DNA ligase?

A
  • joining Okazaki fragments
    • covalent bonds between sugar and phosphate
  • repairs damaged DNA
25
Q

What is the function of topoisomerase?

A
  • helicase unwinds DNA
    • creates tension
  • makes a cut in one strand
    • holds to the spot
    • helix spins (no overwinding)
  • reconnects the spot
  • in E. coli —> DNA gyrase
26
Q

What is a leading and a lagging strand?

A
  • strands in anti-parallel
  • leading strand follows the fork
27
Q

What is the direction of replication?

A
  • from 5’ to 3’
  • starting with sugar ending with phosphate group
28
Q

What is the function of non-coding regions of DNA?

A
  • satellite DNA
    • components of heterochromatin and centromeres
  • telomeres
    • repetitive regions at the end of chromosome
    • protection
  • introns
    • within genes
    • removed in RNA splicing
  • non-coding RNA genes
    • not translated into protein
    • genes for tRNA
  • gene regulatory sequences
    • transcription
    • promoters, enhancers and silencers
29
Q

What are the steps of PCR?

A
  • polymerase chain reaction
  1. denaturation
    • heated to 95ºC
    • separation
  2. annealing
    • cooled to 55ºC
    • primers anneal
  3. elongation
    • 72ºC
    • Taq polymerase
  • the cycle is repeated several time to obtain many copies of DNA
30
Q

What is the role of Taq polymerase in PCR?

A
  • DNA polymerase that is heat-resistant
31
Q

What is DNA sequencing?

A

> unknown DNA in tube + enzymes necessary for replication
dideoxyribonucleotides are added
- fluorescent
- attach to bases but after them no new bases added
fragments separated by length
- electrophoresis
colour and length of fragments compared —> sequence