Lecture 8: PROTEINS IN ACTION - 2 Flashcards

1
Q

What does deoxyhaemoglobin have?

A

A dished haem

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2
Q

What does oxygen in oxyhemoglobin do?

A

Flatten the haem and pulls histidine F8 and helix F towards the binding site (increasing its binding affinity for oxygen)

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3
Q

What weakens oxygen binding?

A

Anything that keeps helix F away

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4
Q

What does the oxygen in oxyhemoglobin do to the fe2+?

A

Moves it into the plane of the haem, draws His F8 down and repositions the helix F

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5
Q

What are shifts in protein secondary elements called?

A

Conformation changes

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6
Q

What is an example of the conformational change?

A

Helix F moving relative to helix c

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7
Q

Where is the most change observed?

A

At the FG loop

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8
Q

What happens to side chain packing in R and T states?

A

The interaction between side chains of the F helix and C helix change

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9
Q

Positions of side chains in R and T states are both …

A

stable

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10
Q

The transition state between R and T is …

A

Unstable so not much time is spent in that position

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11
Q

Describe the changes when going from R to T conformation?

A

The F helix moves so that the interaction between threonine and aspartic acid is borked and an interaction with a different threonine and asparagine is formed. This makes the binding pocket larger

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12
Q

What is BPG?

A

2,3-bisphosphoglycerate

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13
Q

What is BPG?

A

An allosteric inhibitor of oxygen binding to haemoglobin

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14
Q

What is the charge of BPG?

A

Highly negatively charged

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15
Q

How does BPG bind to deoxyhaemoglobin?

A

By electrostatic interaction

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16
Q

What does BPG do?

A

Stabilises haemoglobin in the deoxygenated T state, reducing oxygen affintiy

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17
Q

When is BPG produced?

A

During respiration in peripheral tissues, so promotes oxygen release where it is needed

18
Q

When is cooperativity prominent?

A

Only in presence of allosteric inhibitors of binding

19
Q

What do allosteric inhibitors BPG, CO2 and H+ do?

A

Stabilise the t state which unmasks cooperativity

20
Q

What happens in the absence of inhibitors?

A

Stripped haemoglobin is predominantly in the r state so shows little cooperativity (acts almost as myoglobin)

21
Q

What does elevated CO2, and low pH (elevated H+) do?

A

In metabolising tissues it reduces the affinity of haemoglobin for oxygen, known as the Bohr effect

22
Q

What can CO2 do?

A

Bind to the extreme N terminal amino group

23
Q

What can H+ do?

A

Protonate certain amino acid side chains which contributes to stabilising the deoxyhaemoglobin conformation

24
Q

what is the first substantial adaptation to high altitude?

A

An increase in BPG

25
Q

What does an increase in BPG do?

A

Reduces haemoglobin oxygen binding

26
Q

What does a rightward shift of the curve do?

A

Deliver more oxygen to the tissues

27
Q

What do different amino acid sequences of normal haemoglobin subunits in foetus do?

A

Alter their oxygen binding properties

28
Q

What does foetal haemoglobin include?

A

Alternate isoforms (epsilon, gamma and zeta) which have higher affinities for oxygen

29
Q

What does foetal haemoglobin allow?

A

The foetus to capture oxygen in the placenta

30
Q

What does foetal haemoglobin lack?

A

An amino acid in the BPG binding site so bind BPG less well

31
Q

What is methaemoglobin?

A

A damaged form of haemoglobin where oxidation of haem fe2+ to Fe3+ shifts one subunit to the R state conformation without oxygen bound

32
Q

How does methaemoglobin impair the function?

A

The methaemoglobin does not bind oxygen despite otherwise being in the R state because of the Fe3+.
The subunits of the tetramer are shifted to the R state so don’t release oxygen in the tissues as they should

33
Q

What does the enzyme cytochrome b5 reductase do?

A

regenerates haemoglobin by reducing haemoglobin back to Fe2+ state using transfer of electrons from NADH

34
Q

What are most haemoglobin abnormalities caused by?

A

Genetic mutations

35
Q

What is the mutation in HbM/Boston haemoglobin?

A

His E7 mutation to Try E7 changes the environment causing Fe2+ to oxidize to Fe3+

36
Q

What happens as a result of the mutation in HbM/boston haemoglobin?

A

The haem plane moves slightly, breaking the connection of Fe-HisF8

37
Q

What state is HbM in after the mutation?

A

It remains in the T state, with low affinity for oxygen as the larger tyrosine blocks some of the binding space

38
Q

What does the sick shape do in sickle cell anaemia?

A

Means the red blood cells get stuck in blood capillaries which causes a range of secondary debilitating effects in the person

39
Q

What happens to sickled red blood cells?

A

They become worse at carrying oxygen

40
Q

What is the mutation in sickle cell anaemia?

A

A gain of function (Hb Beta EGV)

41
Q

What does the beta EGV mutation enable?

A

Abnormal hydrophobic interactions between Hb molecules, particularly when in the deoxygenated form, causing polymerisation of Hb into chains the distort the red blood cells