Intro to Macromolecules, protein structure and enzyme catalysis Flashcards
why aren’t lipids true macromolecules?
the monomers aren’t covalently bound
what are sugars?
straight chain polyhydroxy alcohols with an aldehyde (aldose) or ketone (ketose) group
what is glucose?
a 6 carbon sugar with an aldehyde group at carbon 1
how many chiral centres does glucose have?
4 (C2, C3, C4, C5)
what enantiomers of glucose exist?
alpha and beta-ring enantiomers
what is the stable conformation of the glucose ring called?
the ‘chair’ conformation
how many possible chair conformations does beta glucose have?
2
at what position does fructose have a ketone group?
C2
which has fewer branches: starch or glycogen?
starch
what do cell-surface carbohydrates consist of?
a core pentasaccharide with additions
what do nucleotides consist of?
a sugar linked to 3 phosphate residues and a base
at what position is deoxyribose de-oxy?
the 2-position
what sort of sugar is deoxyribose?
an aldopentose
what end of a DNA strand are nucleotides added to?
3’ end
what is the difference between the 3’ end and 5’ end of DNA strands?
5’ end has free phosphate, 3’ end has free hydroxyl
which bases are purine?
adenine and guanine
which bases are pyrimidine?
thymine, uracil and cytosine
how many double stranded regions does tRNA have?
4
what is a ribozyme?
an RNA molecule that acts as an enzyme
what form is the chiral centre of amino acids in proteins?
L form
what form do amino acids exist in at neutral pH
Zwitterion form
why are unbound Cys side chains unusual?
they are often paired as covalent disulphide bonds
why is proline unusual?
side chain is covalently linked to N atom of amino group of amino acid- give rigid cyclic structure
why is the peptide bond a planar structure?
delocalisation of electrons from N onto the bond give it partial double bond characteristics so no rotation around bonds possible and bond length is between single and double (resonance hybrid bond)
what are the bonds in amino acids where rotation is possible?
link points- central C atoms of each residue and their bonds with N and C
what is the phi bond?
bond between central C and N in protein backbone
what is the psi bond?
bond between the central C and carboxyl C in protein backbone
what causes folding of the polypeptide chain?
rotation of phi and psi angles for each residue
what does the Ramachandran plot show?
combinations of angles found in protein structures
what is the alpha helix?
a single right-handed helix
what forms the alpha helix?
C=O oxygen on the peptide bond of 1 residues interacting with N-H hydrogen of residue 4 along the chain
what are 3(10) helices?
right handed helices where the H bonds are between i and i+3
what are the different properties of alpha and 3(10) helices?
alpha helices are less tightly coiled, more stable and more flexible
what is the secondary structure of proteins?
localised regular arrangement of polypeptide backbone formed due to H bonding between N-H and C=O groups of peptide bond
when are H bonds strongest?
when linear
what links secondary structure elements in polypeptide chains?
loop regions
what is a beta sheet?
strands of polypeptide chain that interact with neighbouring strands through N-H to C=O hydrogen bonding forming puckered sheets
which are more flexible: alpha helices or beta sheets?
beta sheets
what is the order of strength of protein interactions?
covalent bonds > H bonds = electrostatic interactions > Van der Waals bonds
what is the tertiary structure?
the overall 3D fold of protein
what determines tertiary structure?
side chains
when do hydrogen bonds form?
when H atom bonded to electronegative atom, so H has partial positive charge
what is the hydrophobic effect?
non-polar residues won’t form H bonds with water molecules, disrupts water-water H bonding so water molecules become ordered around hydrophobic side chains- hydrophobic groups cluster together
what are electrostatic interactions?
strong bonds formed between oppositely charged chemical groups
what is the difference in electrostatic interactions within hydrophobic cores vs in aqueous solution?
in aqueous solution the charges are shielded by interactions between water molecules, in hydrophobic core strength of interactions increases as no water
what are Van der Waals forces?
weak interactions between all atoms, occur as atoms can be transiently polarised by asymmetric electronic distribution
what are disulphide bridges?
disulphide bonds between neighbouring Cys residues as result of oxidation
why don’t disulphide bridges form within the cell cytosol?
they occur due to oxidation so don’t form in intracellular reducing environment
when will a protein stop folding into secondary structures?
when the polypeptide chain reaches a minimum energy conformation determined by maximising formation of these bonds
how many helices form a collagen chain?
3
what is the function of collagen?
provides strength to skin, bone, cartilage and tendons
is the collagen superhelix left or right handed?
left
what allows the helices in the collagen chain to pack together closely?
glycine residues in regular pattern
what causes tight bends in the collagen helices?
proline residues
what is the major driving force for folding of globular proteins?
the hydrophobic effect
how are strands of beta sheets generally depicted?
flat arrows pointing from N terminus to C terminus
how are alpha helices generally depicted?
as cylinders or helices
what are motifs/super-secondary structures?
commonly observed groups of secondary structural elements
what is the Βαβ motif held together by?
H bonds between strands and hydrophobic interactions between helix and strands
what does the βαβ motif often bind to?
DNA
what does an α loop α loop motif usually bind?
divalent cations
what are α helical hairpins held together by?
hydrophobic and ionic interactions
what are β sheets in Greek key motifs held together by?
H bonds between strands
what is a β meander?
antiparallel beta sheets that keep going
why is a proline residue required for a β hairpin turn?
proline has bend to backbone
what is a domain?
part of the protein chain that forms a compact unit with a globular cores, several motifs put together
what are multidomain proteins?
proteins with multiple domains formed from a single polypeptide chain
how many regions does functional insulin have?
2
which regions does functional insulin have?
A and B
what is the structure of preproinsulin?
linear structure, 3 regions (A, B, C)
why is insulin not a quaternary protein?
originally synthesised as a single linear chain
what is a homodimer?
a dimeric protein with 2 of the same polypeptides
what is a heterodimer?
a dimeric protein with 2 different polypeptides
what are antibodies?
proteins used by immune system for molecular recognition
what cells produce antibodies?
B lymphocyte cells
what is the antigen surface an antibody binds to?
the epitope
how many chains do antibodies have?
2 identical light chains, 2 identical heavy chains
what does the secondary structure of antibody heavy and light chains consist of?
4-stranded anti-parallel beta-sheet and 3-stranded anti-parallel beta-sheet stabilised by single disulphide bond
what are the domains in the light chains of antibodies?
1 constant domain and 1 N-terminal variable domain
what are the domains in the heavy chains of antibodies?
3 relatively conserved immunoglobulin domains, 1 N terminal variable domain
what are cofactors?
molecules used by proteins to provide chemical reactivity, not found in a.a. side chains
what will happen if cofactors are removed from the protein?
they are integral to structure so it will unfold
what is a co-substrate?
a molecule that is loosely bound and used once by a protein then released
what is a coenzyme?
a cofactor used in enzyme-catalysed reactions
what is the haem group of myoglobin/haemoglobin an example of?
a prosthetic group
what is a prosthetic group?
cofactor tightly attached to protein and used multiple times, integral to structure of protein
what is the function of myoglobin?
facilitates diffusion of oxygen around muscle tissue
what is myoglobin?
small monomeric protein found in vertebrate muscle
how many residues does myoglobin have?
153
how many alpha-helices does myoglobin have?
8
what is the (Fe)II ion at the centre of the myoglobin haem coordinated by?
4 N atoms, 1 histidine side chain, free 6th position to interact with O2
where is the haem group in myoglobin?
wedged in hydrophobic pocket
what is the function of haemoglobin?
carries oxygen in the blood, binds it in high O2 conc in lungs and releases in respiring tissues
how many subunits does haemoglobin have?
2 alpha-subunits, 2 beta-subunits
what does each subunit of haemoglobin resemble
myoglobin
what is the structure of each subunit of haemoglobin?
8 α-helices, buried haem group, binds oxygen in 6th coordination position
what are the oxygen binding properties of myoglobin?
simple hyperbolic binding curve
what are the oxygen binding properties of haemoglobin?
sigmoidal binding properties, 4 subunits bind cooperatively
how does cooperativity work in haemoglobin?
oxygen binding causes haem ring to adopt straight conformation with Fe(II) moving into ring centre. pulls His residue that coordinates Fe(II) towards oxygen moving α-helix which contains His, places strain on interface between subunits
what is the shared property of the external surface of membrane-spanning proteins?
hydrophobic
what does a hydrophilic internal surface of transmembrane proteins allow for?
recognition and transport of specific hydrophilic molecules
what is the membrane spanning part of most membrane proteins formed from?
alpha helices or beta barrels
how many identical subunits make up the potassium channel?
4
how many alpha helices do each subunit of the potassium channel have?
2
what allows the potassium channel to be embedded in the membrane?
external surface of the protein is hydrophobic
what allows the flow of ions through the potassium ion channel?
hydrophilic pore down centre of tetramer
what does the selectivity filter of the potassium channel do?
provides oxygen atoms from backbone C=O groups so ion can lose hydration water without loss of energy- and therefore can pass through the channel- doesn’t do this for Na+ as Na+ ion is smaller
how many K+ are allowed through the potassium channel for every Na+?
1000
what is the principle of X-ray crystallography?
protein crystals diffract X-rays, diffraction pattern can be used to calculate electron density which can be used to build model of overall 3D structure of protein
what proteins is NMR spectroscopy an option for?
proteins smaller than 300 residues
what happens in cryoelectron microscopy?
samples placed onto a grid and studied at very low temperatures. doesn’t require crystals
what sort of techniques are used to separate a protein of interest based on size or charge?
fractionation techniques
what is the simplest form of chromatography?
affinity chromatography
what happens in affinity chromatography?
column material contains molecule that specifically binds to protein of interest. when sample passed through column and washed only protein of interest binds, others washed away
how can the protein of interest be eluted from the column in affinity chromatography?
by adding competitive ligand
what happens in ion exchange chromatography?
uses column with charged bead material- proteins bind to different degrees depending of charge. protein eluted from column using gradient of increasing cell concentration- salt disrupts electrostatic interactions between protein and column
which proteins elute first in ion exchange?
loosely associated proteins elute first, most tightly associated require greater salt concentration to elute
what is the outcome of gel filtration?
separates proteins by size
what happens in gel filtration?
column resin contains a gel, as sample passes through column proteins can enter gel beads and be slowed by material. larger proteins pass around beads so move faster
what determines structure of a protein?
its amino acid sequence
what determines function of a protein?
its structure
how can structure and function of unknown proteins be predicted?
comparing to known proteins as proteins with same structure often have similar sequences and functions
what are zinc fingers?
simple domains found in DNA/RNA binding protein which can bind to DNA double helix
what is the structure of the zinc finger?
zinc ion at base of finger coordinated by 4 amino acid residues- 2 His and 2 Cys
where do zinc fingers insert into the DNA double helix?
in the major groove
what do misfolded proteins often form?
amyloids- insoluble fibrous aggregates
what diseases are caused by amyloids?
Alzheimer’s and prion diseases
what happens in Alzheimer’s?
amyloids formed from 40-42 residue fragment from membrane protein in brain tissue
what happens in prion diseases?
prion protein misfolds to form toxic filaments, misfolding occurs when PrP protein comes into contact with other misfolded PrP protein
what are Bradford assays used for?
to determine total amount of protein present in a sample
what dye is used in Bradford assays?
Coomassie Brilliant Blue
what colour is Coomassie Brilliant Blue in acidic solution and when bound to protein?
green-brown in acidic solution, blue when bound to protein
what happens in SDS Page?
protein sample heated in presence of SDS, which denatures it and gives it negative charge, sample then undergoes gel electrophoresis to separate proteins by size
what is SDS?
a charged detergent
what does ELISA stand for?
enzyme linked immunosorbent assay
what happens in ELISA?
antibody that binds specifically to protein of interest is linked to solid support. protein will then bind to antibody, other protein molecules washed away. 2nd antibody linked to enzyme binds different site on immobilised protein. washed again. when chemical substrate added enzyme reacts to generate molecule that’s easily detected
what do reactions catalysed by enzymes show?
greater rates, greater specificity (no side reactions), capacity for regulation
how can enzymes make unfavourable reactions take place?
couple them to favourable reaction to provide driving force
what is enthalpy?
heat
why may a reaction need an enzyme?
due to thermodynamic or kinetic barrier
what change in enthalpy is favourable?
negative (release in heat)
what is entropy?
disorder
what change in entropy is favourable?
becoming more disordered
what is the Gibbs free energy equation?
change in enthalpy - temperature(change in entropy)
what change in Gibbs free energy is favourable?
negative
what formula relates equilibrium constant to change in Gibbs free energy?
∆G=RT(ln([B]/[A])- lnK)
what is a favourable reaction, exergonic or endergonic?
exergonic
what is standard free energy?
∆G=RTlnK, free energy for a reaction where starting point is clearly defined and each chemical species is initially present at 1M, pH is 7, temperature is 298K
how do biological systems use ATP in reaction coupling?
hydrolysis of the phosphoanhydride bond of ATP releases energy, if coupled to unfavourable reaction then overall reaction can proceed
how do enzymes reduce the kinetic barrier of reactions?
stabilise transition state, thus decreasing free energy of activation
how do enzymes lower energy of transition state?
acid base catalysis, covalent catalysis, proximity effects, substrate strain
what is the active site?
the location of an enzyme where a reaction takes place
what properties must an enzyme active site have?
correct shape to bind the substrate, correct shape to stabilise transition state, allow product release by not binding product too tightly
what is the lock and key model of enzyme activity?
early model, suggested active site of enzyme is rigid and complementary to substrate, substrate then fits into active site like key in lock
what is the difference between the real model of enzyme binding (induced fit) and the lock and key model?
in real model binding is often flexible, as active site and substrate interact they become deformed- to become more complementary to each other
what reaction is catalysed by carbonic anhydrase?
interconversion of CO2 + H2O <-> HCO3- + H+
what does the active site of carbonic anhydrase contain?
an essential Zn ion coordinated in 3 positions by His side chains with a free 4th coordination position, a binding pocket for a CO2 molecule
how do carbonic anhydrases work?
substrate water molecule occupies 4th coordination position of Zn ion, binding to Zn deforms the water, causes electron movement towards O atom. this polarisation weakens bonds within the water. another His residue is placed close to the substrate water and accepts a proton from it leaving OH- ion sttached to zinc. binding pocket for CO2 brings CO2 molecule into close proximity with OH-, electron rich O of OH- attacks positive carbon of CO2 resulting in formation of bicarbonate
what are proteases?
enzymes that cut protein molecules by hydrolysis of peptide bonds
what are examples of serine proteases?
thrombin, elastase, trypsin, chymotrypsin
what is in the active site of serine proteases?
essential catalytic serine residue, charge relay system (His and Asp residues), specificity pocket, oxyanion hole
what is the function of thrombin?
blood clotting
what is the function of elastase, trypsin and chymotrypsin?
digestive enzymes
what makes up the charge relay system in serine proteases?
a Ser residue, His residue and Asp residue which make up a catalytic triad
what is the first stage in the reaction mechanism of serine proteases?
substrate binding: catalytic serine residue attacks carbonyl carbon of peptide bond forming covalent bond between substrate and enzyme. positive charge stabilised by His residue which is then stabilised by Asp residue
what determines the substrate specificity of a serine protease?
specificity pocket, side chain of substrate peptide preceding peptide bond fits into specificity pocket
what is a charge relay mechanism?
charges that develop during catalysis are stabilised by presence of neighbouring opposite charges
what sort of peptides does trypsin select?
ones with positively charged Lys/Arg in R1 as these fit in specificity pocket
what sort of peptides does chymotrypsin select?
ones with bulky hydrophobic residues (Phe, Trp, Tyr) in R1 as has large selectivity pocket
what allows the active site of serine proteases to transiently stabilise the tetrahedral intermediate?
oxyanion hole
what is the oxyanion hole?
a region of the active site of serine proteases which is well suited to bind to and stabilise a negatively charged oxygen ion through H bonding to backbone NH groups of Gly and Ser
why does the oxyanion hole stabilise the intermediate and not the substrate in the serine protease mechanism?
carbonyl oxygen of substrate isn’t positioned to correctly enter the hole but formation of the tetrahedral intermediate distorts it placing negative oxygen in oxyanion hole
what happens after the serine protease oxyanion hole stabilises the tetrahedral intermediate?
intermediate decomposes due to proton donation from His57, releases half of substrate as new peptide and remains attached to other 1/2 as acyl-enzyme intermediate
what happens in stage 2 of the serine protease reaction mechanism?
active site regenerated, removing part of original substrate. water molecule enters active site, acts as nucleophile attacking carbonyl carbon of substrate peptide. charge relay and oxyanion hole stabilise 2nd tetrahedral intermediate. second part of substrate peptide released, enzyme regenerated
what is the function of Asp proteases?
cut peptide bonds
what is the difference between the mechanism of serine proteases and Asp proteases?
Asp proteases use pair of catalytic Asp side chains instead of reactive serine residue
how do Asp proteases work?
one Asp (Asp1) begins reactive cycle in protonated state- donates proton to substrate to form transition state. Asp2 deprotonated at start of reaction cycle, accepts proton to stabilise positive charge in transition state (acts as base). Asp2 removes proton from water, activated water performs nucleophilic attack on C=O at peptide bond to be broken. C-N bond broken, N takes proton from Asp1
how many people were living with AIDS in 2021?
around 38 million
how many people died from AIDS in 2021?
over 500,000
what are polyproteins?
multiple proteins linked covalently in inactive form
what is the function of HIV protease?
cuts polyproteins to generate functional HIV molecules
what sort of protein is HIV protease
an Asp protease
what are ritonavir and saquinavir?
drugs which act as competitive inhibitors for HIV protease
where does HIV protease cleave peptide bonds?
between bulky residues
where is the active site of HIV protease?
at interface of the 2 chains that make up the molecule
what are class 1 enzymes?
oxidoreductases
what are oxidoreductases?
enzymes that catalyse oxidation and reduction
what is a common type of oxidoreductase?
a dehydrogenase
what is a dehydrogenase?
oxidoreductase which catalyses removal of electrons with a proton
what is an alcohol dehydrogenase?
enzyme which removes hydrogen from alcohols forming aldehyde
what are class 2 enzymes?
transferases
what is a transferase?
enzyme which transfers a functional group from a donor molecule to an acceptor molecule
what is a protein kinase?
a transferase that transfers a phosphate group onto a protein molecule
what are class 3 enzymes?
hydrolases
what is a hydrolase?
enzyme that catalyses hydrolytic cleavage reactions, breaking molecules by severing covalent bonds
what is a protease?
a hydrolase which cuts a protein
what is a DNase?
a hydrolase which cuts DNA
what are class 4 enzymes?
lyases
what is a lyase?
an enzyme which breaks covalent bonds by means other than hydrolysis or oxidation and creates new double bonds or rings in the process
what is aldolase?
a lyase which breaks fructose-1,6-bisp to make 2 smaller molecules, generates C=O in the process
what are class 5 enzymes?
isomerases
what are isomerases?
enzymes which catalyse geometric changes- alter relative positions of chemical groups on molecule without altering groups present
what are class 6 enzymes?
ligases
what are ligases?
enzymes which join molecules together using ATP
what do enzyme kinetics do?
analyse rate at which enzyme works and how rate varies depending on concentration of substrate + presence of regulatory molecules
what are the uses of enzyme kinetics?
info about mechanism of action of enzyme, effect of different molecules on kinetics shows how enzyme is controlled in cell, quantify effectiveness of drugs, info about function in tissue
what does hexokinase do?
converts glucose into glucose-6-phosphate, allows GluTs to take up glucose
what does the different kinetics of glucokinase and hexokinase allow?
muscle to continue to take up glucose when blood sugar levels drop, liver to secrete glucose under these conditions
what is the simplest form of enzyme kinetics?
Michaelis-Menten kinetics
what does Michaelis-Menten enzymes do?
combine with substrate S to form non-covalent enzyme substrate complex which decomposes to form product
what do Michaelis-Menten reactions assume?
there is no conversion of product back into substrate- only true during early stages of reaction when concentration of product very low so initial rates of reactions always measured
what do rate constants do?
relate rate of reactions (v) to substrate concentration
how is rate of production of product determined by k2 (rate constant for conversion of enzyme substrate complex to product) in Michaelis-Menten kinetics?
V=k2[ES]
what relationship does the Michaelis-Menten equation describe?
relationship between initial rate of a reaction (v0) and initial substrate concentrations [S]
what does derivation of the Michaelis-Menten equation assume?
fixed concentration of enzyme, fixed reaction conditions, enzyme conc. small compared to initial substrate conc
what is the Michaelis-Menten equation?
v0= Vmax[S]/Km+[S]
what does the relationship between V0 and [S] follow in Michaelis-Menten kinetics?
rectangular hyperbola
what equation relates Vmax to enzyme concentration?
Vmax=k2[E]T
what is [E]T?
total enzyme concentration for fixed enzyme concentration
what is Vmax?
the maximum possible reaction rate, achieved when enzyme supplied with infinite substrate concentration
what is Km?
substrate concentration when rate of reaction is half maximum rate
what equation gives Km?
Km= (k-1 +k2)/k1
what will the Km of an enzyme with a high affinity for its substrate be?
very low
what is the gradient of a plot of 1/Vo against 1/[S]?
Km/Vmax
what is the x-intercept of a plot of 1/Vo against 1/[S]?
-1/Km
what is the y-intercept of a plot of 1/Vo against 1/[S]?
1/Vmax
what do competitive inhibitors do?
bind directly to enzyme active site, compete with substrate for access- limit the time during which active site accessible to substrate so reduce rate of reaction
what make the best competitive inhibtors?
transition state analogues, as active site optimised to bind to transition state
what properties make HIV protease inhibitors good competitive inhibitors?
tetrahedral C residue attached to OH to mimic tetrahedral intermediate, lack N atom of peptide bond- prevents hydrolysis by the protease- so binds tightly and can’t be cleaved
what is the effect of a competitive inhibitor on Vmax and Km?
Vmax doesn’t change, Km will increase as greater conc of substrate needed to reach half-maximal rate
what is Vmax?
rate of reaction at infinite substrate concentration
what is the effect of a non-competitive inhibitor on Vmax and Km?
won’t change Km, will decrease Vmax
what do non-competitive inhibitors do?
alter conformation of important catalytic residue without altering amino acids that bind to the substrate
what do allosteric inhibitors do?
alter shape of enzyme so active site less complementary to reaction transition state
what is the effect of allosteric inhibitors on Vmax and Km?
decrease Vmax, increase Km
how do nerve gases work?
irreversibly react by forming covalent bond with essential serine residue in active site of acetylcholine esterase which breaks down ACh- so muscle remains stimulated when nerve gases inhibit the enzyme- prevents breathing
how does suicide inhibition work?
inhibitor resembles substrate but can’t go through complete enzyme-catalysed reaction. covalently binds to enzyme so enzyme can’t bind actual substrate- irreversible
how does penicillin work?
suicide inhibitor. inhibits glycopeptide transpeptidase- essential for formation of peptide bonds in synthesis of bacterial cell wall
why do suicide inhibitors make ideal drug molecules for targeting enzymes from pathogens?
have high specificity and are irreversible
what is the most common form of reversible covalent modification at the active site?
addition of a phosphate group to enzyme
which residues can be phosphorylated in the active site?
Ser, Tyr, Thr
how does phosphorylation of the active site inhibit enzyme activity?
bulky phosphate group blocks substrate access
how is Cdk2 controlled by phosphorylation?
phosphorylation of Tyr15 in active site of Cdk2 blocks binding of ATP into active site, prevents function of enzyme. removed when cell ready to divide
how are zymogens activated?
part of N-terminal region cut away, so protein can undergo conformational change and active site can adopt high activity form
what additional site do allosteric enzymes have?
site distinct from active site where allosteric effectors can bind
how do allosteric activators work?
alter conformation of enzyme so active site is in better conformation to bind to/stabilise transition state
what do mutations in glucokinase cause?
reduce activity of enzyme, reduce responsiveness of pancreas to glucose and thus insulin secretion
how do allosteric activators work on glucokinase?
bind to site 20A away from active site, stabilise high energy conformation of glucokinase increasing its glucose sensitivity (decreases Km) and increasing rate of conversion to G-6-P (increases Vmax)
what is the simplest case of sigmoidal kinetics?
binding of substrate causes enzyme to undergo conformational change from low activity conformation to high
what is homotropic allostery?
when the substrate binding causes the conformational change
what does homotropic allostery do?
facilitates control of enzyme function- small change in substrate conc can lead to large change in enzyme activity
what is heterotropic allostery?
when molecules in addition to the substrate cause conformational changes to determine enzyme activity
what are the kinetics of an allosteric enzyme at high activator concentration so all subunits in high activity form?
standard hyperbolic kinetics
what happens to enzyme kinetics when allosteric inhibitor added to allosteric enzyme?
remain sigmoidal but higher substrate concentrations required for high activity
what is phosphofructokinase (PFK)?
tetrameric enzyme in glycolytic pathway
what reaction does PFK catalyse?
conversion of fructose-6-phosphate to frucose-1,6-bisphosphate
effect of ATP of PFK pathway?
highly active when ATP concs low and reduced flux when ATP levels high
what allosterically inhibits PFK?
ATP
what allosterically activates PFK?
AMP
what property is shown by the 4 subunits of PFK?
cooperativity
what stabilises the high activity state of PFK?
fructose-6-phosphate- the substrate
what reaction does glycogen phosphorylase catalyse?
first stage of breakdown of glycogen- forming glucose-1-phosphate
what pathway does glucose-1-phosphate enter?
glycolysis
properties of glycogen phosphorylase?
dimeric enzyme, show cooperativity
difference between high and low activity forms of glycogen phosphorylase?
in high activity form active site is exposed to protein surface, in low active site is buried and inaccessible to substrate
what allosterically inhibits glycogen phosphorylase?
ATP and glucose-6-phosphate (when these are abundant, glycogen breakdown not needed)
what allosterically activates glycogen phosphorylase?
AMP
effect of phosphorylation of glycogen phosphorylase?
activates enzyme stabilising high activity form.
how is the phosphorylated and dephosphorylated form of glycogen phosphorylase differentiated?
glycogen phosphorylase a= phosphorylated, glycogen phosphorylase= dephosphorylated
target of Tamiflu and Relenza?
influenza neuraminidase- catalyses reaction needed for release of mature virus proteins
structure of Tamiflu to make it good inhibitor of influenza neuraminidase?
transition state analogue with additional chemical groups to occupy empty active site pockets + interact with active site side chains, positively charged group on 4 position to fill pocket in active site + form H bonds with 2 glutamate residues in this pocket, positively charged group made smaller, oxygen in ring exchanged for C to increase stability
