1. Enzymes

2. Definition of an enzyme
Enzyme is protein catalyst (i.e. increase the rate of reactions) NOT changed during the reaction Enzymes direct all metabolic reactions occurring in the cells

3. Classification of enzymes
1- Oxidoreductase: catalyses oxidation-reduction reactions 2-Transferases: catalyses transfer of C-, N- or P- containing groups 3-Hydrolases: catalyses breakdown of bonds by addition of water (hydrolysis) 4- Lyases: catalyses breakdown of C – C , C-S & C-N bonds 5- Isomerase: catalyses racemization of isomers (D- to L- or L- to D-) 6- Ligases: catalyses formation of bonds between carbon and O, S, N

6. Mechanism of action of enzymes: lock & key model+
E S ES complex E P
An enzyme binds a substrate in a region called the active site
Enzyme active site is rigid (stable, not malleable)
Only certain substrate can fit the active site of the enzyme
(i.e. the enzyme can act on only one or few substrates. i.e. specific) P S S P

7. Mechanism of enzyme action: induced fit model
Active site of the enzyme adjust s its shape to bind the substrate.
Enzyme structure is flexible (not rigid)
This mechanism gives increased range of substrate specificity (many types of substrates can bind the same enzyme)
E S ES complex E P P S S S P

8. Properties of enzymes
1- All Enzymes are Proteins (except ribozymes that are RNA)
2- Active Site
Each enzyme molecules contain a special pocket called the active site.
The substrate bind with the active site to form enzyme substrate-complex
(ES) which is converted to enzyme-product complex (EP).
EP dissociates to enzyme and product
3-Catalytic Efficiency:
Each enzyme molecule is capable of transforming 100 – 1000 substrate
molecules into product each second

9. Properties of enzymes (cont.)
4- Specificity: Enzymes are highly specific i.e. each enzyme interacts with one or few type of substrates and catalyzes one type of reactions 5- Holoenzymes: Some enzymes require molecules other than proteins for enzymic activity. Holoenzymes: apoenzyme (protein part) + nonprotein component Apoenzymes are inactive without the nonprotein component non-protein part: 1- a metal ion as zinc or ferrous ions (called cofactors) 2- small organic molecules (called coenzymes) Coenzymes : may be only transiently associated with the enzyme or may be permanently associated with the enzyme (prosthetic group) Coenzymes may be derived from vitamins. (e.g. NAD+ contains niacin)

10. assistance

11. Properties of enzymes (cont.)
6- Regulation: Enzyme activity can be regulated by activation or inhibition. 7- Location within the cell: Many enzymes are localized in specific organelles within the cell. This serves to isolate the reaction substrate or product from other opposite reactions.

12. Factors affecting enzyme activity: 1- Temperature
Optimum temperature
Reaction
Rate
Low High
Temperature
1- Little activity at low temperature
2- Rate of activity increases with temperature
3- Most active at optimum temperatures
(usually 37°C in all enzymes of humans)
4- Activity is lost with denaturation at high
temperatures

13. Factors affecting enzyme activity: 2- pH
Reaction
Rate pH
Maximum activity at optimum pH
where:
R groups of amino acids have proper charge
Tertiary structure of enzyme is intact
Most enzymes lose activity in very low or high pH (due to loss of tertiary structure i.e. denatured)
Each enzyme has its own optimum pH
Optimum pH
Each enzyme has its own optimum pH

14. Each enzyme has its own optimum pH
level of blood

15. Factors affecting enzyme activity: 3- Substrate concentration
Maximum Activity
Reaction
Rate
Substrate Concentration
Increasing substrate concentration increases the rate of reaction (with enzyme concentration is constant)
Maximum activity reached when all of enzyme molecules combine with substrate

16. Factors affecting enzyme activity: 4- Enzyme concentration
Reaction
Rate
Enzyme Concentration
Increasing enzyme concentration increases the rate of reaction (with substrate concentration is constant)

17. Factors affecting enzyme activity: 5- Inhibition of enzyme activity
Inhibitor:
is an any substance that can diminish the velocity of a reaction which is catalyzed by an enzyme.
Reversible Inhibitors: bind to enzymes through non-covalent bonds
Irreversible Inhibitor: bind to enzymes through covalent bonds
Types of inhibitors according to site of binding of inhibitor:
Competitive Inhibitor : inhibitor binds at the active site of the enzyme
Noncompetitive Inhibitor: inhibitor binds to a site other than active site

18. Km of enzyme
Km (Michaelis Constant) of an enzyme is numerically equal to the substrate concentration at which the velocity of reaction is equal to 1/2 Vmax
Km is the substrate concentration at which 1/2 maximal velocity is reached
If Km is small,
the substrate concentration required for the reaction to reach 1/2 maximal velocity is small.
i.e. the enzyme has a high affinity for the substrate.
If Km is large,
the substrate concentration required for the reaction to reach
1/2 maximal velocity is large.
i.e. the enzyme has a low affinity for the substrate

20. Competitive inhibitors
A competitive inhibitor:
has a structure similar to substrate of the enzyme
occupies active site of the enzyme
competes with substrate for occupying the active site of the enzyme
increases Km of the enzyme to its substrate.
i.e. more substrate is required to reach ½ Vmax
effect can be reversed by increasing substrate concentration.

21. Reversible Inhibition:Competitive

22. Noncompetitive inhibitors
A noncompetitive inhibitor:
Has a structure different from the substrate
Binds to the enzyme at a site different from active site
i.e. does not compete with the substrate of the enzyme for the active site of the enzyme
Changes the shape of enzyme and thus active site shape is changed.
Accordingly, substrate cannot fit in the altered active site
So, no reaction occurs
Effect is not reversed by adding more substrate
Does not change Km of the enzyme (i.e. increase of substrate concentration will not lead to reaching of ½ Vmax).

24. Regulation of enzyme activity
The regulation of the activity of enzyme is essential for coordinating the metabolic processes.
Types of regulation:
1- General: (occurs in all types of enzymes in the body)
increasing substrate concentration will lead to increase activity of the
enzyme
2-Special regulatory mechanisms: (not all enzymes of the body)
i- Allosteric effectors
ii- Covalent modification
iii. Increase or decrease rate of enzyme synthesis

25. 1- Allosteric effectors
Allosteric binding sites are sites on the enzyme different from the active site.
Binding of an allosteric effectors to this site will make changes in shape of the whole enzyme with an effect on activity.
So, the activity may be decreased (positive allosteric effector) or
decreased (negative allosteric effector).

26. 2- Covalent modification
1- Many enzymes may be regulated by addition of phosphate groups to the enzyme (modification by phosphorylation) Addition of phosohate group may cause activation or inactivation of the enzyme 2- Some enzymes are released as an inactive form (zymogen) By removal of a part of the enzyme (modification), it will be active.

28. 3- Increasing or decreasing rate of enzyme synthesis
By this mechanism cells regulate the amount of enzyme by changing the rate of enzyme synthesis
The increase (induction) or decrease (repression) of enzyme synthesis leads to change total amount of active site.
This mechanism is slow (takes from hours or days)

29. Medical importance of enzymes of blood
Blood enzymes can be classified into two major groups:
1- Enzymes that have functions in blood (a smaller group):
They are present in blood in high amounts
Example: liver secretes zymogens (inactive precursors) involved in
blood coagulation.
2-Enzymes that do not have functions in blood (a large number)
These enzymes are released from cells during normal cell turn over.
They have functions in cells (intracellular)
BUT: they do not have a function in blood
In healthy individuals, levels of these enzymes are constant .
So, the presence of elevated enzyme level in blood may indicate tissue
damage that leads to increase in release of these enzymes.

30. Medical importance of enzymes of blood (cont.)
Many diseases that cause tissue damage results in increased release of intracellular enzymes into plasma (blood)
So, the enzyme levels in blood are measured for diagnosis of these diseases
Diseases of the heart, liver, skeletal muscles and other tissues are diagnosed by an elevation of a blood enzymes.
The level of elevation of an enzyme correlates with the extent of tissue damage in any of these organs.
Some enzymes may be available in high amount in only one organ
So, the elevation of blood levels of these enzymes are diagnostic for
diseases of this organ only. (specific)
Example: Alanine aminotransferase (ALT) enzyme elevation in blood
indicates disease of the liver (specific for liver cells).