Semester 2 Flashcards
High / Low Affinity Binding Curve:
What does the release of the FIRST oxygen do?
It causes a conformational change in the other 3 subunits.
It converts them from a relaxed state to a more tense state. (Back to a lower affinity!)
So we have EASIER release of the 2nd oxygen, which makes it even more tense!
And then the same process with the 3rd and 4th oxygen…
High / Low affinity binding curve:
Why is this type of curve NOT seen in Myoglobin?
Due to its structure: It only has 1 subunit and therefore NO cooperative activity.
Identify the correct characteristic of Hydroxyproline + Hydroxylysine
They are required for proper functioning of Collagen.
The protein capsid of a virus has what structure?
Primary, secondary, tertiary AND quaternary structure!
Which compound is an inhibitor of Nitric Oxide Synthase?
N-Methylarginine
Methyllysine has the following important roles…
Epigenetics + Myosin function
Food Pickling affects protein structure by disrupting…
Electrostatic Interactions
Which amino acid is commonly found in Beta Turns?
Cis- Proline
Which of the following cofactors is required for the formation of Gamma- Carboxyglutamate?
Vitamin K
The effect of pH on the binding of O2 to Haemoglobin is known as the…
Bohr Effect
The following type of Alpha-Helix is commonly found in membranes to form Pores…
Amphipathic
An elite athlete trains for 4 weeks at 4,500m. What is the efficiency of O2 transport from the lungs to the tissues?
(Graph)
37%
What molecule provides proteins with ‘Elastic’ properties?
Desmosine
What amino acid is a ‘classic’ disruptor of Alpha- Helices?
Proline
What is an INCORRECT characteristic for protein alpha- helices?
A channel is formed down the centre of the alpha helix.
Phosphoserine, phosphothreonine and phosphotyrosine are formed by…
Kinases
What methods can be used to determine the 3D structure of proteins?
X-Ray Crystallography or Nuclear Magnetic Resonance (NMR)
What roles do metals perform in Restriction Enzymes?
They help ALIGN the Scissile bond correctly in the active site and they REDUCE electrostatic repulsion.
The Aconitase enzyme converts Citrate to Isocitrate through…
Sequential processes of Dehydration/ Hydration
What roles does Mg2+ play in the functioning of Hexokinases?
It REDUCES electrostatic repulsion and correctly orientates the SUBSTRATES of the hexokinase reaction.
Why is Vitamin D currently a ‘hot topic’?
It is thought to reduce infection by the Covid-19 virus.
How can UV-visible Spectroscopy be used to detect oxygen levels in the blood?
By measuring the RATIO of the Soret band absorbance at 414nm and 429nm.
What is the biochemical role of Glutathione Peroxidase?
It converts Hydrogen Peroxide to water, utilising GSH as a reductant!
What is the biochemical / physiological function of Carbonic Anhydrase?
It is an enzyme that REVERSIBLY converts C02 to carbonic acid OR carbonic acid to C02.
What are the functions of the Cytochrome P450 enzymes?
Vitamin D Synthesis
Testosterone and Oestrogen Synthesis
Bile Salt Synthesis
Drug Metabolism
What is the function of bile salts?
They act as Biological Detergents in the small intestine and facilitate excretion of EXCESS Cholesterol.
What amino acid binds to the Iron-Sulfur centre of Aconitase?
Cysteine
An enzyme has a Km of 0.5mM while the substrate concentration is 10mM.
At what % of Vmax is the enzyme catalysing the conversion of substrate to product?
V0 = Vmax x [S] divided by KM + [S]
(Michaelis - Menten Equation)
For KM = 0.5mM
and
[S] = 10mM
( If it’s a percentage of Vmax then assume the Vmax is always 100%!)
V0 = 100% x 10 divided by (0.5 + 10)
V0 = 95% of Vmax!
An enzyme has a Km of 0.5mM, a Vmax of 100 mmoles of product formed per minute and the substrate concentration is 10mM.
Calculate V0.
V0 = Vmax x [S] divided by KM + [S]
(Michaelis - Menten Equation)
KM = 0.5 Vmax = 100mmoles product per min [S] = 10mM
V0 = 100 x 10 divided by 0.5 + 10
V0 = 95 mmoles product formed per min!
(V0 is ALWAYS IN THE SAME UNIT as Vmax!)
The reaction catalysed by Hexokinase is as follows: Glucose +ATP > Glucose-6-phosphate + ADP
What is the role of ATP in this reaction?
Provide energy to drive the reaction forward AND to donate a phosphate (Pi)
What parameters are affected by doubling enzyme concentration?
Vmax and Vi
What compound binds PERMANENTLY to a protein?
Prosthetic Group
What molecule is NOT an enzyme cofactor?
Glutamate
Enzymes are biological catalysts that…
INCREASE the rate of reaction by REDUCING the Transition State Energy Barrier
The affinity of an enzyme for a substrate is indicated by the…
Km
The Km of a reaction is 5mM, Vmax is 20nmoles / minute and substrate concentration is 5nM.
What is the rate of reaction Vi? (Interchangeable with V0)
V0 = vmax x [S] divided by km + [S]
V0 = 20 x 5 divided by 5 + 5
V0 = 10 nmoles / min (SAME UNITS AS VMAX!)
What mechanims of Rate Enhancement are used by Chymotrypsin?
Acid base catalysis
Covalent catalysis
Electrostatic catalysis / environment
Proximity + orientation effects
B with a lone pair of electrons in covalent and base catalysis is a…
Base Catalyst
NADPH donates … electrons to Cytochrome P450 Reductase, which donates… electrons to Cytochrome P450
2
1
In the catalytic mechanism of Chymotrypsin, His 57 acts as a…
BOTH base and acid catalyst
What rate enhancement mechanism is NOT used by Chymotrypsin?
Metal Ion Catalysis
Base Catalysis is defined as…
Temporary proton abstraction
In Elastase, the catalytic triad consists of…
Asp-102, Ser-195 and His-57
The substrate specificity of Trypsin is defined by a binding pocket that contains…
A negatively charged Aspartate residue
The catalytic mechanism of chymotrypsin, trysin and elastase is a…
2 substrate ping pong mechanism
What is INCORRECT regarding covalent catalysis?
Cannot take place with hydroxyl groups
The first step in catalysis by Chymotrypsin is…
Substrate binding
Rate Enhancement strain and distortion takes place in…
Lysozyme and Endonucleases
If you add 200ul of your 6M ethanol to the reaction, what is the FINAL concentration in the cuvette during the reaction? Final volume = 2.4 ml.
Vi x Ci = Vf x Cf
Vi = 200ul Vf = 2400ul (CONVERT UNITS) Ci = 6000mM (as answer was in mM, if theres a M and an mM value, convert to mM!)
Cf = 200 x 6000 / 2400
= 500mM
In the alcohol dehydrogenase practical (ADH), you measured the initial rates (Vi) of the following reaction:
C2H5OH + NAD+ > CH3CHO + NADH + H+
Which component is DIRECTLY measured?
NADH increase (you are measuring the change in absorbance at 340nm ie. the conversion of NAD+ to NADH)
In the ADH practical you measured the initial rate over … seconds
30
You are using a P200 pipette set at 050. What volume will this measure?
50 ul
In order to measure the REDUCTION of NAD+ to NADH your spectrophotometer wavelength will be set to…
340 nm
In the ADH practical, the spectrophotometer should be ‘blanked’ by…
Checking the correct wavelength has been set, inserting a cuvette containing 2.5ml water OR buffer, then pressing ‘measure blank’
In order to measure the OXIDATION of NADH to NAD+ your spectrophotometer wavelength will be set to…
340nm
You have measured the absorbance of 1nM solution of pure compound Y at 340nm.
The pathlength of the cuvette is 1cm. If the measured absorbance is 0.05, what is the extinction coefficient of compound Y?
Beere-Lambert equation: Abs =ecl
E = Abs/ cl
= 0.05 / (1mM x1cm )
= 50 mM -1 cm -1
Which component of the enzyme reaction is added immediately before oressing ‘measure sample’ on spectrophotometer?
Enzyme
What is NOT a common function of carbohydrates?
Catalysing reactions in cells
Which carbon atom in a glucose ring can have a hydroxyl group bound in an ‘Up’ or ‘Down’ orientation?
Anomeric carbon
Which bond links glucose monomers in Maltose?
Alpha 1-4 glycosidic bonds
Which is NOT a storage carbohydrate in plants?
Glycogen
What is FALSE about cellulose?
Coils are stabilised by hydrogen bonds
Mushroom cell walls are made from…
Chitin
Which enzyme is most likely to hydrolyse a Beta 1-4 glycosidic bond?
Endochitinase
What property of cellulose makes it HARD for most mammals to digest?
The anomeric state of its monomers!