13.14: Enzymatic Catalysis

Learning Objectives

• To describe the interaction between an enzyme and its substrate.

 

Enzyme-catalyzed reactions occur in at least two steps. In the first step, an enzyme molecule(E) and the substrate molecule or molecules (S) collide and react to form an intermediate compound called the enzyme-substrate (E–S) complex. (This step is reversible because the complex can break apart into the original substrate or substrates and the free enzyme.) Once the E–S complex forms, the enzyme is able to catalyze the formation of product (P), which is then released from the enzyme surface:

 

S+E→E–S (13.14.1)

E–S→P+E (13.14.2)

 

Hydrogen bonding and other electrostatic interactions hold the enzyme and substrate together in the complex. The structural features or functional groups on the enzyme that participate in these interactions are located in a cleft or pocket on the enzyme surface. This pocket, where the enzyme combines with the substrate and transforms the substrate to product is called the active site of the enzyme (Figure 13.14.3).

Figure 13.14.3: Substrate Binding to the Active Site of an Enzyme. The enzyme dihydrofolate reductase is shown with one of its substrates: NADP⁺ (a) unbound and (b) bound. The NADP⁺ (shown in red) binds to a pocket that is complementary to it in shape and ionic properties.

The active site of an enzyme possesses a unique conformation (including correctly positioned bonding groups) that is complementary to the structure of the substrate, so that the enzyme and substrate molecules fit together in much the same manner as a key fits into a tumbler lock. In fact, an early model describing the formation of the enzyme-substrate complex was called the  lock-and-key model (Figure 13.14.4). This model portrayed the enzyme as conformationally rigid and able to bond only to  substrates that exactly fit the active site.

Figure 13.14.4: The Lock-and-Key Model of Enzyme Action. (a) Because the substrate and the active site of the enzyme have complementary structures and bonding groups, they fit together as a key fits a lock. (b) The catalytic reaction occurs while the two are bonded together in the enzyme-substrate complex.

Working out the precise three-dimensional structures of numerous enzymes has enabled chemists to refine the original lock-and-key model of enzyme actions. They discovered that the binding of a substrate often leads to a large conformational change in the enzyme, as well as to changes in the structure of the substrate or substrates. The current theory, known as the induced-fit model, says that enzymes can undergo a change in conformation when they bind substrate molecules, and the active site has a shape complementary to that of the substrate only after the substrate is bound, as shown for hexokinase in Figure 13.14.5. After catalysis, the enzyme resumes its original structure.

Figure 13.14.5: The Induced-Fit Model of Enzyme Action. (a) The enzyme hexokinase without its substrate (glucose, shown in red) is bound to the active site. (b) The enzyme conformation changes dramatically when the substrate binds to it, resulting in additional interactions between hexokinase and glucose.

The structural changes that occur when an enzyme and a substrate join together bring specific parts of a substrate into alignment with specific parts of the enzyme’s active site. Amino acid side chains in or near the binding site can then act as acid or base catalysts, provide binding sites for the transfer of functional groups from one substrate to another or aid in the rearrangement of a substrate. The participating amino acids, which are usually widely separated in the primary sequence of the protein, are brought close together in the active site as a result of the folding and bending of the polypeptide chain or chains when the protein acquires its tertiary and quaternary structure. Binding to enzymes brings reactants close to each other and aligns them properly, which has the same effect as increasing the concentration of the reacting compounds.

Example \(\PageIndex{1}\)

1. What type of interaction would occur between an OH group present on a substrate molecule and a functional group in the active site of an enzyme?
2. Suggest an amino acid whose side chain might be in the active site of an enzyme and form the type of interaction you just identified.

Solution

1. An OH group would most likely engage in hydrogen bonding with an appropriate functional group present in the active site of an enzyme.
2. Several amino acid side chains would be able to engage in hydrogen bonding with an OH group. One example would be asparagine, which has an amide functional group.

 

Exercise \(\PageIndex{1}\)

1. What type of interaction would occur between each group present on a substrate molecule and a functional group of the active site in an enzyme?

   1. COOH
   2. NH₃⁺
   3. OH
   4. CH(CH₃)₂

2. What type of interaction would occur between each group present on a substrate molecule and a functional group of the active site in an enzyme?

   1. SH
   2. NH₂
   3. C₆H₅
   4. COO⁻

3. For each functional group in Exercise 1, suggest an amino acid whose side chain might be in the active site of an enzyme and form the type of interaction you identified.

Answer

Question 1: 1. hydrogen bonding 2. ionic bonding 3. hydrogen bonding 4. dispersion forces

Question 2: 1. disulfide bonding  2.  ionic bonding 3. dispersion forces 4.  hydrogen bonding 

Question 3:

1. The amino acid has a polar side chain capable of engaging in hydrogen bonding; serine (answers will vary).
2. The amino acid has a negatively charged side chain; aspartic acid (answers will vary).
3. The amino acid has a polar side chain capable of engaging in hydrogen bonding; asparagine (answers will vary).
4. The amino acid has a nonpolar side chain; isoleucine (answers will vary)

One characteristic that distinguishes an enzyme from all other types of catalysts is its substrate specificity. An inorganic acid such as sulfuric acid can be used to increase the reaction rates of many different reactions, such as the hydrolysis of disaccharides, polysaccharides, lipids, and proteins, with complete impartiality. In contrast, enzymes are much more specific. Some enzymes act on a single substrate, while other enzymes act on any of a group of related molecules containing a similar functional group or chemical bond. Some enzymes even distinguish between D- and L-stereoisomers, binding one stereoisomer but not the other. Urease, for example, is an enzyme that catalyzes the hydrolysis of a single substrate—urea—but not the closely related compounds methyl urea, thiourea, or biuret. The enzyme carboxypeptidase, on the other hand, is far less specific. It catalyzes the removal of nearly any amino acid from the carboxyl end of any peptide or protein.

 

                                                             

Enzyme specificity results from the uniqueness of the active site in each different enzyme because of the identity, charge, and spatial orientation of the functional groups located there. It regulates cell chemistry so that the proper reactions occur in the proper place at the proper time. Clearly, it is crucial to the proper functioning of the living cell.

Summary

A substrate binds to a specific region on an enzyme known as the active site, where the substrate can be converted to product. The substrate binds to the enzyme primarily through hydrogen bonding and other electrostatic interactions. The induced-fit model says that an enzyme can undergo a conformational change when binding a substrate. Enzymes exhibit varying degrees of substrate specificity.