6.3: Proteins

Learning Objectives

By the end of this section, you will be able to do the following:

1. Describe some examples of how protein functions
2. Discuss the relationship between amino acids and proteins
3. Describe how amino acids are classified
4. Explain the four levels of protein organization
5. Describe the ways in which protein shape and function are linked

Proteins are one of the most abundant organic molecules in living systems and have the most diverse range of functions of all macromolecules. Proteins may be structural, regulatory, contractile, or protective. They may serve in transport, storage, or membranes; or they may be toxins or enzymes. Each cell in a living system may contain thousands of proteins, each with a unique function. Their structures, like their functions, vary greatly. They are all, however, amino acid polymers arranged in a linear sequence.

Types and Functions of Proteins

Enzymes, which living cells produce, are catalysts in biochemical reactions (like digestion) and are usually complex or conjugated proteins. Each enzyme is specific for the substrate (a reactant that binds to an enzyme) upon which it acts. The enzyme may help in breakdown, rearrangement, or synthesis reactions. We call enzymes that break down their substrates catabolic enzymes. Those that build more complex molecules from their substrates are anabolic enzymes, and enzymes that affect the rate of reaction are catalytic enzymes. Note that all enzymes increase the reaction rate and, therefore, are organic catalysts. An example of an enzyme is salivary amylase, which hydrolyzes its substrate amylose, a component of starch.

Hormones are chemical-signaling molecules, usually small proteins or steroids, secreted by endocrine cells that act to control or regulate specific physiological processes, including growth, development, metabolism, and reproduction. For example, insulin is a protein hormone that helps regulate the blood glucose level. Table \(\PageIndex{1}\) lists the primary types and functions of proteins.

Protein Types and Functions

Type	Examples	Functions
Digestive Enzymes	Amylase, lipase, pepsin, trypsin	Help in food by catabolizing nutrients into monomeric units
Transport	Hemoglobin, albumin	Carry substances in the blood or lymph throughout the body
Structural	Actin, tubulin, keratin	Construct different structures, like the cytoskeleton
Hormones	Insulin, thyroxine	Coordinate different body systems' activity
Defense	Immunoglobulins	Protect the body from foreign pathogens
Contractile	Actin, myosin	Effect muscle contraction
Storage	Legume storage proteins, egg white (albumin)	Provide nourishment in early embryo development and the seedling

Table \(\PageIndex{1}\): Examples of the various functions a protein may have.

Proteins have different shapes and molecular weights. Some proteins are globular in shape; whereas, others are fibrous in nature. For example, hemoglobin is a globular protein, but collagen, located in our skin, is a fibrous protein. Protein shape is critical to its function, and many different types of chemical bonds maintain this shape. Changes in temperature, pH, and exposure to chemicals may lead to permanent changes in the protein's shape, leading to loss of function, or denaturation. Different arrangements of the same 20 types of amino acids comprise all proteins. Two rare new amino acids were discovered recently (selenocystein and pirrolysine), and additional new discoveries may be added to the list.

Amino Acids

Amino acids are the monomers that comprise proteins. Each amino acid has the same fundamental structure, which consists of a central carbon atom, or the alpha (α) carbon, bonded to an amino group (NH₂), a carboxyl group (COOH), a hydrogen atom and an R group (Figure \(\PageIndex{1}\)). The R group varies between amino acids and is also referred to as the side chain.

Figure \(\PageIndex{1}\): Amino acids have a central asymmetric carbon to which an amino group, a carboxyl group, a hydrogen atom, and a side chain (R group) are attached.

Scientists use the name "amino acid" because these acids contain both amino group and carboxyl-acid-group in their basic structure. As mentioned, there are 20 common amino acids present in proteins. Nine of these are essential amino acids in humans because the human body cannot produce them and we obtain them from our diet. For each amino acid, the R group (or side chain) is different (Figure \(\PageIndex{1}\)).

With the exception of glycine (which has a single hydrogen as an R group), each amino acid can exist in two enantiomeric forms, because the central carbon is a chiral center. This difference may seem small to our eyes (Figure \(\PageIndex{2}\)), but in the cell, only L-amino acids are used to make proteins.

Figure \(\PageIndex{2}\): The central carbon in an amino acid is a chiral carbon (bonded to four unique groups) and thus can exist in two enantiomeric forms. Shown here is L- and D- alanine. (Image by Synpath, CC BY-SA 4.0)

The chemical nature of the side chain determines the amino acid's nature (that is, whether it is acidic, basic, polar, or nonpolar). For example, the amino acid glycine has a hydrogen atom as the R group. Amino acids such as valine, methionine, and alanine are nonpolar or hydrophobic in nature, while amino acids such as serine, threonine, and cysteine are polar and have hydrophilic side chains. The side chains of lysine and arginine are positively charged, and therefore these amino acids are also basic amino acids. Proline has an R group that is linked to the amino group, forming a ring-like structure. Proline is an exception to the amino acid's standard structure since its amino group is not separate from the side chain (Figure \(\PageIndex{3}\)).

A three-letter abbreviation or a single letter represents amino acids. For example, the letter V or the three-letter symbol val represent valine.

Visual Connection

Figure \(\PageIndex{3}\): There are 20 common amino acids commonly found in proteins, each with a different R group (variant group) that determines its chemical nature.

Give it a try! You don't need to memorize the amino acid's names or structures for this course, but you should be able to determine which group they belong to based on the chemical characteristics of their R groups.

Peptide Bonds

The sequence and the number of amino acids ultimately determine the protein's shape, size, and function. A peptide bond is formed between amino acids to create a polymer, through a dehydration reaction. One amino acid's carboxyl group and the incoming amino acid's amino group combine, releasing a water molecule. The resulting bond is the peptide bond (Figure \(\PageIndex{4}\)).

Figure \(\PageIndex{4}\): Peptide bond formation is a dehydration synthesis reaction. The carboxyl group of one amino acid is linked to the incoming amino acid's amino group. In the process, it releases a water molecule.

The products that such linkages form are peptides. As more amino acids join to this growing chain, the resulting chain is a polypeptide. Each polypeptide has a free amino group at one end. This end is known as the N terminal, or the amino terminal, and the other end has a free carboxyl group, also the C or carboxyl terminal. While the terms polypeptide and protein are sometimes used interchangeably, a polypeptide is technically a polymer of amino acids, whereas the term protein is used for a polypeptide or polypeptides that have combined together, often have bound non-peptide prosthetic groups, have a distinct shape, and have a unique function. After protein synthesis (translation), most proteins are modified. These are known as post-translational modifications. They may undergo cleavage, phosphorylation, or may require adding other chemical groups. Only after these modifications is the protein completely functional.

Protein Structure

As we discussed earlier, a protein's shape is critical to its function. For example, an enzyme can bind to a specific substrate at an active site. If this active site is altered because of local changes or changes in overall protein structure, the enzyme may be unable to bind to the substrate. To understand how the protein gets its final shape or conformation, we need to understand the four levels of protein structure: primary, secondary, tertiary, and quaternary.

Primary Structure

Amino acids' unique sequence in a polypeptide chain is its primary structure. For example, the pancreatic hormone insulin has two polypeptide chains, A and B, and they are linked together by disulfide bonds. The N terminal amino acid of the A chain is glycine; whereas, the C terminal amino acid is asparagine (Figure \(\PageIndex{5}\)). The amino acid sequences in the A and B chains are unique to insulin.

Figure \(\PageIndex{5}\): Bovine serum insulin is a protein hormone comprised of two peptide chains, A (21 amino acids long) and B (30 amino acids long). In each chain, three-letter abbreviations that represent the amino acids' names in the order they are present indicate primary structure. The amino acid cysteine (cys) has a sulfhydryl (SH) group as a side chain. Two sulfhydryl groups can react in the presence of oxygen to form a disulfide (S-S) bond. Two disulfide bonds connect the A and B chains together, and a third helps the A chain fold into the correct shape. Note that all disulfide bonds are the same length, but we have drawn them different sizes for clarity.

The gene encoding the protein ultimately determines the unique sequence for every protein. A change in nucleotide sequence of the gene’s coding region may lead to adding a different amino acid to the growing polypeptide chain, causing a change in protein structure and function. In sickle cell anemia, the hemoglobin β chain (a small portion of which we show in Figure \(\PageIndex{6}\)) has a single amino acid substitution, causing a change in protein structure and function. Specifically, valine in the β chain substitutes the amino acid glutamic. What is most remarkable to consider is that a hemoglobin molecule is comprised of two alpha and two beta chains that each consist of about 150 amino acids. The molecule, therefore, has about 600 amino acids. The structural difference between a normal hemoglobin molecule and a sickle cell molecule—which dramatically decreases life expectancy—is a single amino acid of the 600. What is even more remarkable is that three nucleotides each encode those 600 amino acids, and a single base change (point mutation), 1 in 1800 bases causes the mutation.

Figure \(\PageIndex{6}\) Because of this change of one amino acid in the chain, hemoglobin molecules form long fibers that distort the biconcave, or disc-shaped, red blood cells and causes them to assume a crescent or “sickle” shape, which clogs blood vessels (Figure \(\PageIndex{7}\)). .his 3.4 • Proteins 81 The beta (β)- chain of hemoglobin is 147 amino acids in length, yet a single amino acid substitution in the primary sequence leads changes in secondary, tertiary and quaternary structures and sickle cell anemia. In normal hemoglobin, the amino acid at position six is glutamate. In sickle cell hemoglobin glutamate is replaced by valine. Credit: Rao, A., Tag, A. Ryan, K. and Fletcher, S. Department of Biology, Texas A&M University.

Because of this change of one amino acid in the chain, hemoglobin molecules form long fibers that distort the biconcave, or disc-shaped, red blood cells and causes them to assume a crescent or “sickle” shape, which clogs blood vessels (Figure \(\PageIndex{7}\)). This can lead to myriad serious health problems such as breathlessness, dizziness, headaches, and abdominal pain for those affected by this disease. William Warrick Cardozo showed that sickle-cell anemia is an inherited disorder, meaning that the difference in the specific gene's encoding region is passed down from parents to children. As you will learn in the genetics unit, the inheritance of such traits is determined by a combination of genes from both parents, and these very small differences can have significant impacts on organisms.

Figure \(\PageIndex{7}\) In this blood smear, visualized at 535x magnification using bright field microscopy, sickle cells are crescent shaped, while normal cells are disc-shaped. (credit: modification of work by Ed Uthman; scale-bar data from Matt Russell)

Secondary Structure

The local folding of the polypeptide in some regions gives rise to the secondary structure of the protein. The most common are the α-helix and β-pleated sheet structures (Figure \(\PageIndex{8}\)). Both structures are held in shape by hydrogen bonds. 

Figure \(\PageIndex{8}\) The α-helix and β-pleated sheet are secondary protein structures formed when hydrogen bonds form between the carbonyl oxygen and the amino hydrogen in the peptide backbone. Certain amino acids have a propensity to form an α-helix while others favor β-pleated sheet formation. Black = carbon, White = hydrogen, Blue = nitrogen, and Red = oxygen. Credit: Rao, A., Ryan, K. Fletcher, S. and Tag, A. Department of Biology, Texas A&M University.

In a α-helix , The hydrogen bonds form between the oxygen atom in the carbonyl group in one amino acid and another amino acid that is four amino acids farther along the chain. Every helical turn in an alpha helix has 3.6 amino acid residues. The polypeptide's R groups (the variant groups) protrude out from the α-helix chain. In the β-pleated sheet, hydrogen bonding between atoms on the polypeptide chain's backbone form the "pleats". The R groups are attached to the carbons and extend above and below the pleat's folds. The pleated segments align parallel or antiparallel to each other, and hydrogen bonds form between the partially positive hydrogen atom in the amino group and the partially negative oxygen atom in the peptide backbone's carbonyl group. The α-helix and β-pleated sheet structures are in most globular and fibrous proteins and they play an important structural role.

Tertiary Structure

The polypeptide's unique three-dimensional structure is its tertiary structure (Figure \(\PageIndex{9}\)). This structure is in part due to chemical interactions at work on the polypeptide chain. Primarily, the interactions among R groups create the protein's complex three-dimensional tertiary structure. The nature of the R groups in the amino acids involved can counteract forming the hydrogen bonds we described for standard secondary structures. For example, R groups with like charges repel each other and those with unlike charges are attracted to each other (ionic bonds). When protein folding takes place, the nonpolar amino acids' hydrophobic R groups lie in the protein's interior; whereas, the hydrophilic R groups lie on the outside. Scientists also call the former interaction types hydrophobic interactions. Interaction between cysteine side chains forms disulfide linkages in the presence of oxygen, the only covalent bond that forms during protein folding.

Figure \(\PageIndex{9}\) A variety of chemical interactions determine the proteins' tertiary structure. These include hydrophobic interactions, ionic bonding, hydrogen bonding, and disulfide linkages.

All of these interactions, weak and strong, determine the protein's final three-dimensional shape. When a protein loses its three-dimensional shape, it may no longer be functional.

Quaternary Structure

In nature, some proteins form from several polypeptides, or subunits, and the interaction of these subunits forms the quaternary structure. Weak interactions between the subunits help to stabilize the overall structure. For example, insulin (a globular protein) has a combination of hydrogen and disulfide bonds that cause it to mostly clump into a ball shape. Insulin starts out as a single polypeptide and loses some internal sequences in the presence of post-translational modification after forming the disulfide linkages that hold the remaining chains together. Silk (a fibrous protein), however, has a β-pleated sheet structure that is the result of hydrogen bonding between different chains.

Protein Structure Summary

Figure \(\PageIndex{10}\) summarizes the four levels of protein structure (primary, secondary, tertiary, and quaternary). 

Figure \(\PageIndex{10}\) Observe the four levels of protein structure in these illustrations. Credit: Rao, A. Ryan, K. and Tag, A. Department of Biology, Texas A&M University.

Sort the definitions below to the appropriate level of protein structure.

Video Summary

The video below reviews the structure of an amino acid and the levels of protein structure. Since proteins are so complex and diverse, it's also important to think about how we, as scientists, represent protein structures. Perhaps not surprisingly, there is more than one way to do this, each with its own strengths and weaknesses. Choosing which to use depends on the purpose of your diagram.

Denaturation and Protein Folding

Each protein has its own unique sequence and shape that chemical interactions hold together. If the protein is subject to changes in temperature, pH, or exposure to chemicals, the protein structure may change, losing its shape without losing its primary sequence in what scientists call denaturation. Denaturation is often reversible because the polypeptide's primary structure is conserved in the process if the denaturing agent is removed, allowing the protein to resume its function. Sometimes denaturation is irreversible, leading to loss of function. One example of irreversible protein denaturation is frying an egg. The albumin protein in the liquid egg white denatures when placed in a hot pan. Not all proteins denature at high temperatures. For instance, bacteria that survive in hot springs have proteins that function at temperatures close to boiling. The stomach is also very acidic, has a low pH, and denatures proteins as part of the digestion process; however, the stomach's digestive enzymes retain their activity under these conditions.

Protein folding is critical to its function. Scientists originally thought that the proteins themselves were responsible for the folding process. Only recently researchers discovered that often they receive assistance in the folding process from protein helpers, or chaperones (or chaperonins) that associate with the target protein during the folding process. They act by preventing polypeptide aggregation that comprise the complete protein structure, and they disassociate from the protein once the target protein is folded.

Link to Learning

Complete the Protein Folding Interactive by slowing scrolling down the page. This interactive reviews the structure of an amino acid, classification of amino acids and the levels of protein structure discussed above.