Enzyme regulation

Enzymes – regulation

Allosteric – Regular binds non-active site and activates/inhibits/inactivates enzymes (sometimes a

downstream product in negative feedback)

- Type of non-competitive inhibition

o Not in the active site of the enzyme – induce a change in conformation (inhibit to
catalyze)

Covalent (reversible) – Phosphate or other group activates/inhibits/inactivates enzymes

o Can also be seen in methyl grps etc.

o Phosporylation
o Reversible --attachment

Enzyme itself -Zymogen aka proenzyme is irreversible

- Pepsinogen (zymogen) encounter HCl, cleave, turns to active form pepsin

- Activated by cleavage.
- -ogen ending
- Released in inactive, then cleave, then irreversibly activated (useful)

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 Biocatalyst – increase rates of reactions

 Regulation – activation and inhibition
 Regulation and metabolism
o Feedback inhibition
o Allosteric regulation
o Phosphorylation
o Molecules that aid enzymes

Feedback inhibition

- Product of a pathway will inhibit an enzyme

 Allosteric regulation

- Allos – other; Steric-Object

- Change in shape to have regulation
Ex: Inhibition
- Allosteric site instead of active site

- Can also be for activation

Phosphorylation – inactivate or activate enzyme

- Bind phosphate
- Muscles: Epinephrine (adrenaline)

Molecule “Partners” of enzyme

- Cofactors – inorganic ions that bind to certain enzymes

o Iron (Fe2+, Fe3+)
o Copper (Cu+ or Cu2+)
o Zinc (Zn 2+)
- Coenzymes Carbon-containing molecules, smaller than enzymes
o Biotin
o Coenzyme A
o NAD
o FAD
- Prosthetic groups – permanently bound to their enzymes
o Heme (oxygen carrying protein in blood)
o Flavin

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 o Retinal

Activation

 “Partner” molecules
 Phosphorylation
 Allosteric regulation

Inhibition

 Competitive

- Active feedback inhibition -product of reaction will compete for the active site and bind,
prevent for other reactants to bind and proceeding with the reaction.

 Non-competitive
- Passive feedback inhibition – product bind to allosteric site, switch it off
- Allosteric
- Phosphorylation

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ENZYME INHIBITION

Enzyme inhibition

Enzyme activity can be inhibited. Enzyme inhibitors can be classified according to three different
mechanisms:

- irreversible inhibitors

- competitive inhibitors

- non-competitive inhibitors.

Agents that bind covalently to enzymes and disrupt their function are irreversible inhibitors. A few do
bind non-covalently, and they are highly toxic.

Competitive inhibitors

Compete with the substrate for the active site by binding reversibly with non-covalent bonds at the
active site. They block the substrate from binding to the active site, and forming the enzyme-substrate
complex. A comparative inhibitor will increase the Km, but the Vmax does not change. The inhibition can

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be overcome by adding more substrate. The classic indication that something is a competitive inhibitor
is that the Vmax is the same.

E+S = ES  E+P

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EI

Non-Competitive Inhibitors

Bind non-covalently to a site other than the active site and change the conformation of the enzyme.
They do not prevent the substrate from binding to the enzyme. Unlike competitive inhibitors, they
cannot be overcome by the addition of substrate. The non-competitive inhibitor will also lower the
Vmax due to inability of the reaction to proceed as efficiently. They do not lower the enzymes for the
substrate, so the Km remains the same.

Uncompetitive Inhibition

The inhibitor binds only to the enzyme-substrate complex. This type of inhibition decreases the Vmax
due to less availability of the active complex. The Km will decrease due to better binding efficiently.

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2 Types of Enzyme inhibition

Competitive

Competes with substrate to bind Active Site

-increase substrate to aid this

Noncompetitive

Allosterically binds enzyme elsewhere from active site to inhibit/disable enzymes

Michaelis menten plot

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Higher Km, lower affinity

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Enzymes

- Catalyst, faster
- Proteins, made up amino acids
- Substrate to active site (specificity)
- Enzyme-substrate complex
- Induced fit model
- Optimal range and temperature
- Inhibitors: competitive and non-competitive

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Reversible

-Competitive

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Adding more substrate can add to the probability that it will bind

-Non-competitive

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Lac Operons

Operons – a form of gene control or repression in prokaryotes

Lac operon – a well – understood operon codes for B-galactosidase to break down lactose in E-coli.

RNA polymerase bind to promoter

Operator – if it can reach to gene

- Repressor (protein) join in operator

- Unable to pass through

Lactose

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Repressor -diff conformation and change shape, unable to repress

Free to move

Gene control

8min

Jacob-monod model

Based on your results, what can you conclude about the affinity of the substrate, H2O2?

According to the results in Lineweaver-Burke Plot, Enzyme B will more likely bind to the substrate H202
than Enzyme A. It is due to the Km value being lower in Enzyme B, specifically with the value of 1.28,
than the Km in Enzyme A, with 3.11. With this, the lower Km value indicates higher affinity. Moreover,
the Vmax value of Enzyme B is lower with 0.36, indicating a slower second step compared to Enzyme A
which has the Vmax value of 1.29, since more time and therefore energy is needed to cleave the non-
covalent enzyme-substrate complex.

Of enzymatic activity

Non-covalent, weaker, affinity

since more time and therefore energy is needed to cleave the non-covalent enzyme-substrate complex.

Of enzymatic activity

Non-covalent, weaker, affinity

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