Introduction

Have you ever thought about why athletes or gymnasts are often on a protein-rich diet? It’s
because these people expend more energy compared to people who don’t work out. And a diet
rich in proteins fuels the body to build muscle, promote quick recovery, boost immunity,
replenish glycogen, and burn fat, all of which are important during strenuous workouts. [1]

But are they only required for people who work out? No, protein has tons of roles to play in all
living organisms. It’s one of the essential macronutrients synthesized by organisms required for
a healthy life.

This article is all about what proteins are, their types, and how they are different from one
another. It also covers the functions of proteins in living organisms and methods to study these
molecules in lab conditions.

What Are Proteins?

Proteins are one of the versatile macromolecules in living organisms that serve crucial
functions in different biological processes.

They are present throughout the body of organisms — in muscles, bones, skin, hair, and
virtually every other body part or tissue. [2] They are made of 20 amino acids, arranged in
different structural forms, forming around 10,000 proteins (or even more than that). [2]

Some key properties of proteins responsible for their functions are: [3]

Are proteins linear polymers built of monomer units called amino acids?

Proteins are constructed linearly from 20 amino acids, which is a recurring process in a cell.
But the linear/primary sequence of the protein is not responsible for their functional roles. [3]

The sequences spontaneously fold into different arrangements, forming three-dimensional

structures that are determined by the amino acids present in the sequence. And these folded
structures facilitate the functions of these proteins.[3]

Do proteins contain a wide range of functional groups?

The most predominant functional groups present in proteins are alcohols, thiols, thioethers,
carboxylic acids, carboxamides, and a variety of basic groups.[3]

These functional groups combined in different random patterns in the amino acid sequence
accounts for the broad spectrum of protein function. In simple words, the properties of the
functional groups of proteins determine their enzymatic and other metabolic functions in the
body.[3]

Can proteins interact with one another and other biological macromolecules, forming complex
assemblies?

Proteins interact with other proteins or biomolecules to perform tasks that they alone aren’t
equipped for. Examples of such tasks are DNA replication, the intracellular transmission of
signals, and other complex essential biological processes.[3]

Some proteins are rigid, and some show limited flexibility.

The rigid proteins (have tightly compact structures which restrict any motion or movement) are
involved in the structural formation of cells, such as the cytoskeleton which forms internal
scaffolding within cells.[3]

Whereas flexible proteins (have loosely bound structures that can easily rearrange or move
when required) act as hinges, springs, and levers to perform essential functions and form
complexes with other proteins and macromolecules.[3]

Synthesis of Protein

1. Biological Synthesis
The process of protein synthesis is called translation. In this process, the mRNA codes are
translated into the respective amino acids that are involved in protein formation. Each amino
acid has its unique nucleotide sequence of genes. The genetic code of one amino acid consists
of three sets of nucleotides called codons.[4]

The process starts with the transcription of DNA to pre-mRNA using RNA polymerase. The pre-
mRNA is then modified through post-transcription to form a mature mRNA. [4] The mature
mRNA is used as a template by ribosomes for protein synthesis.

Ribosomes bind to the mRNA and use one codon (consisting of three nucleotides) matching the
anticodon present on tRNA (transfer ribonucleic acid is an adaptor molecule that helps in
decoding mRNA) for the synthesis of one amino acid. [4] The four nucleotides, A (adenine), G
(guanine), C (cytosine), and T (thymine) form a different combination of nucleotides to form
different amino acids.

Figure: An illustration of protein synthesis.

2. Chemical Synthesis

The chemical synthesis of proteins involves peptide synthesis, which uses strategies like
chemical ligation, Staudinger ligation, or other orthogonal chemical reactions to couple
synthetic peptides.[4] Here, peptides (a chain of 30-50 amino acids) are produced and linked
together via amide or peptide bonds to form specific proteins.

But it’s an inefficient technique when it comes to producing a polypeptide chain of more than
300 amino acids. It’s important to note that the chemical synthesis of protein occurs from C-
terminus to N-terminus, whereas in biological processes, the synthesis occurs in the opposite
direction, that is from N-terminus to C-terminus. [4]

Structures of Proteins

Proteins are built by linking two or more amino acid residues together in different orientations
or configurations. The amino acids in the proteins are joined together by peptide bonds formed
through a condensation reaction between two amino acids, releasing water molecules in the
process.

The description of protein structure may sound similar to peptides, but there is a fine line
between proteins and peptides as both differ in size, structure, and functions, as explained
below:

 Peptides are composed of 2 to 50 amino acids and can be classified further into
oligopeptides and polypeptides.[5] The polypeptide is a chain of 20 or more amino
acids, joined together by peptide bonds, while oligopeptides are composed of 20-30
amino acid residues. However, proteins are made up of a long chain of 50 or more
amino acids linked through peptide bonds.
 Peptides mainly have a linear structure, whereas proteins have four structural
levels: primary (linear), secondary, tertiary, and quaternary. [5]
 Peptides are involved in regulating the activities of other molecules while proteins
have a wider spectrum of functions which include structural determination,
enzymatic reactions, and hormonal functions.[5]
Four Levels of Protein Structure
The different levels of protein structure are possible as a result of different chemical
interactions within its structure. For example, the folding of the linear sequence of amino acids
into three-dimensional structures is driven by several non-covalent interactions such as
hydrogen bonding, ionic interactions, Van der Waals forces, and hydrophobic interactions.

Given below is a short description of the four levels of protein structure. [6]
1. Primary Structure

The primary structure is the arrangement of amino acids into a linear polypeptide chain. Here,
the amino acids are only joined by peptide bonds and disulfide bonds. The sequence of amino
acids in a polypeptide chain is responsible for the functions of the proteins, which is decided by
decoding the genetic codes or genes corresponding to the proteins.[6]

Figure: The representation of the primary structure of the protein.[8]

2. Secondary Structure

Secondary structure is the regular, recurring arrangements of adjacent amino acids in a

polypeptide chain.[7] This occurs by forming hydrogen bonds between molecules and it
represents the initial stage of polypeptides folding into three-dimensional structural form, that
is, before tertiary and quaternary structures.

The most common secondary structures are alpha-helix and beta-sheet.

 Alpha helix: In this structural form, the carbonyl group (C=O) of one amino acid is
hydrogen-bonded to the amino hydrogen (N-H) of the fourth amino acid in the
sequence. This pulls the linear chain into a helical ribbon form, with each helical
turn containing 3.6 amino acids.[7]
 Beta sheet or strand: Here, two or more segments of a polypeptide chain are lined
up next to each other. The strands are hydrogen-bonded together in either parallel
or antiparallel forms.[7]

Figure: Structural forms of alpha-helix and beta-pleated sheets.[8]

3. Tertiary Structure
Here, the polypeptide chain further folds into a three-dimensional space, involving one or
several domains. The alpha-helix and beta sheets are folded into a compact globular structure
that’s driven by non-specific hydrophobic interactions.

The interactions occur between polar, nonpolar, acidic, and basic R groups within the
polypeptide chain.[7] When proteins lose the tertiary structural form, they can’t perform any of
their functions.

In an aqueous environment, the hydrophobic or nonpolar groups move into the interior of the
proteins while the hydrophilic R groups lie on the surface or outside the structure. [7]

Figure: The representation of the tertiary structure of the protein.[8]

4. Quaternary Structure

More than one polypeptide chain is involved in the formation of a quaternary structure. The
polypeptide chains can be identical multiple copies or different in amino acid sequences. It’s
the weak interactions such as hydrogen bonding and London dispersion forces, that keep the
multiple polypeptides bound together in forming a quaternary structure. [6]

A well-known example of a quaternary structure is hemoglobin that carries oxygen in the blood
and consists of four peptide chains (two alpha and two beta chains) to form a tetramer. [6]

Figure: An illustration of the four levels of protein structure.[7]

Physicochemical Properties of Protein
1. Color and taste: Proteins are colorless and tasteless. They are homogenous and
crystalline.
2. Molecular weights of proteins: This can be calculated if the molecular weight of
amino acids in a linear chain is known. So the total number of amino acids
multiplied by 110 (subtracting the number of water molecules released during
 condensation reaction) gives the exact molecular weight of proteins. Note: The
average molecular weight of amino acids is 110 Daltons.[9]
3. Coagulation: This is the heat denaturation of proteins like albumins and globulins
to form insoluble coagulates or aggregates, known as coagulum.[9]
4. Solubility: Increase in the acidity or alkalinity of the solution increase the
solubility of the protein. That is, the solubility of proteins depends on the pH of the
solution, and at the isoelectric pH, they are the least soluble. [9]
5. Optical activity: The optical activity of proteins is due to the asymmetry of the
polypeptides chains and at the α-carbon atoms in the amino acid residues. [10] The
proteins rotate the plane for polarized light to the left (they are levorotatory).
6. Denaturation: A protein is said to be denatured when heat, acids, alkalies,
alcohol, acetone, urea, beta-mercaptoethanol cause partial or complete unfolding of
the native conformation of polypeptide chains.[9]
7. Isoelectric pH (pI): The pH at which the protein structure contains an equal
number of positive and negative charges on their functional groups is called the
isoelectric pH. This property is applied in labs to isolate proteins from different
samples. When proteins are subjected to an electric field at isoelectric pH, they
neither move to anode or cathode and become less soluble – this causes
precipitation of the proteins.[9]
8. Electrochemistry of proteins: The electrochemistry of proteins is defined as the
redox potential of proteins, due to the presence of free positive and negative
charges on the functional groups of the amino acids. This electrochemical
character in proteins is mainly decided by the charged ammonium groups (―NH3+)
of lysine and arginine, and negatively charged carboxyl groups (―COO−) of aspartic
acid and glutamic acid. The charges on these side groups of amino acid of proteins
make them move towards respective (cathode or anode) electrodes in the presence
of an electric field and participate in any chemical reaction.[11]
9. Condensation Reaction: Condensation reaction occurs between the carboxyl
group of one amino acid and the amino group of another amino acid. It leads to the
formation of peptide bonds and the release of water molecules.
Classification of Proteinss

A. Based on solubility
1. Fibrous proteins: These are insoluble in water and are mainly involved in
supportive and protective functions in organisms. They are tough, strong, and
linear in shape.[9] Here, long, parallel polypeptide chains are cross-linked together,
forming the protein. Collagen, keratin, silk, and fibrin are common examples of
fibrous proteins.[9]
2. Globular proteins: These are soluble in water and have metabolic functional roles,
such as forming enzymes, hormones, and antibodies. Here, the polypeptide chains
are tightly folded into a sphere. The majority of proteins in the cells belong to this
category. Examples include DNA polymerase, RNA polymerase, and hemoglobin. [9]

B. Based on structural complexity

1. Simple proteins: They have a simple structural organization and are only
composed of amino acid residues. They are also known as homoproteins. They can
be globular or fibrous proteins. Examples include keratin, elastin, albumin,
collagen, and histones.[9]
2. Conjugated proteins: These are complex proteins and here, proteins are loosely
bound with one or more non-protein moieties. The non-protein groups are called
prosthetic groups and they can be carbohydrates, lipids, metal ions, nucleic acid,
phosphoric acid, and FAD.[9] These proteins are usually globular in shape and are
soluble in water. Examples include nucleoprotein, metalloprotein, and lipoproteins.
[9]

3. Derived Proteins: They are low molecular weight derivatives of proteins — derived
from the partial hydrolysis of simple or conjugated proteins by acid, enzyme, or
alkalies. Examples are coagulated proteins, proteans, peptones, and peptides.[9]

Biological Functions of Protein

Proteins perform several essential life-sustaining metabolic functions in organisms. Based on

their functions, they are classified into:[12]

1. Structural proteins: Most of the structural proteins are fibrous proteins and are
insoluble in water. They form the components of bone, tendons, cartilage, skin,
 connective tissue, hair, and horn. Examples include collagen, keratin, and elastin.
[12]

2. Enzymes: They are biological catalysts and work by reducing the activation energy
of reactants. During this process, they speed up the metabolic reactions of cells.
Most of the enzymes are globular conjugated proteins. Examples include
nitrogenase, DNA polymerase, and lipase.[12]
3. Hormones: The protein hormones in the cells include glucagon, insulin, and
adrenocorticotropic hormone.[12]
4. Respiratory pigments: These are colored proteins that are conjugated and contain
pigments (chrome) as their prosthetic group. Examples are hemoglobin and
myoglobin.[12]
5. Contractile proteins: These proteins are involved in muscle contraction at the
expense of energy from ATP molecules. Examples are actin and myosin.[12]
6. Storage proteins: These proteins store metal or amino acids in the cells and are
found in seeds, eggs, milk, and pulses. Examples are casein, gluten, and ferritin. [12]
7. Transport proteins: These proteins are responsible for transporting molecules or
materials to their target destination. They form channels in the plasma membrane
and are also involved in the formation of blood and lymph in animals. Examples
include serum albumin.[12]
8. Defense proteins: These are proteins that are involved in protecting the organism
from foreign microbes or materials. Examples include immunoglobulin (antibodies)
and fibrinogen.[12]
9. Toxins: Snake venoms are toxic proteins.
Methods to Study Proteins
Different characteristics of proteins are studied using different techniques which include: in
vivo, in vitro, and in silico.[4]
In vivo techniques are used to study the functional roles of proteins inside cells; in
vitro techniques are preferred when it comes to understanding the working mechanism of
proteins in a certain environment; while in silico is a computation method of studying
proteins, like understanding protein complex formation and structural determination. [4]

Given below are techniques based on the studies they are used for:[4]

1. Protein purification: The process starts with lysing cells; here, the cell membrane
is disrupted to release all cellular components which are then purified using
ultracentrifugation. It fractionates the cellular components in different fractions
that contain soluble proteins. The proteins in the lysates are precipitated by salting
out and isolated using different chromatography techniques based on molecular
weight, net charge, and binding affinity. The protein is then purified by using either
gel electrophoresis, spectroscopy, or electrofocusing, depending on the experiment
requirements.[4]
2. Presence of protein in samples: Two tests are used to study the presence of
proteins in the given sample: Biuret Test and Ninhydrin Test. In the Biuret test, 2
ml of the sample is added to 2 ml 10% NaOH and one drop of 10% CuSO4 solution.
If the color of the solution changes to violet, it indicates the presence of peptide
linkage.[13] In the ninhydrin test, the formation of violet color after 1 ml of
Ninhydrin solution is added to 1 ml protein solution and gently heated indicates
the presence of α-amino acids in the solution.[13]
3. Structure prediction and determination: Homology modeling is a computational
tool used to predict the protein structure. It’s useful for protein engineering, such
as in designing novel folds for drug applications. [4] X-ray crystallography, NMR
spectroscopy, dual-polarization interferometry, and circular dichroism are other
popular techniques used for the determination of protein structure.[4]
4. Proteomics: Total proteins present in a cell is called proteomes. And the study of
isolation, separation, identification, characterization, and purification of these
proteins is known as proteomics. The most common techniques used for this
purpose are 2D electrophoresis, mass spectrometry, protein microarrays, and two-
hybrid screening.[4]

Conclusion

Proteins are one of the essential biomolecules required to sustain life. They consist of amino
acids arranged in four structural levels: primary, secondary, tertiary, and quaternary.
They differ from peptides at both structural and functional levels. The functions of proteins
range from transportation of molecules, structural formation, to storage and enzymatic roles,
whereas peptides only influence the activities of other molecules.

The spectrum of functions of proteins in organisms has pulled the scientist’s brain towards the
complexity of their actions, hence scientists are curious to understand the structure and
working mechanism of the molecule. Some common tools used to study proteins are mass
spectrometry, chromatography, circular dichroism, and spectrometry.

Working with proteins requires a high level of precaution and expertise. And considering its
importance in organisms, scientists are trying to develop, synthesize, and purify the molecule
at an individual level for application in medical areas. The area of proteomics also opens the
doors for young scientists to make a breakthrough and contribute to enhancing the quality of
human life.
Lipids are fatty compounds that perform a variety of functions in your body. They’re part of
your cell membranes and help control what goes in and out of your cells. They help with
moving and storing energy, absorbing vitamins and making hormones. Having too much of
some lipids is harmful. A lipid panel can tell you if you have normal levels.

What are lipids?

Lipids are chemical compounds (elements that make a chemical bond) in your body that help
with some of its functions. These are fatty or waxy substances your body makes that don’t
dissolve in water.

Examples of lipids

Cholesterol is a lipid in your blood. Your body needs it to help you take in fats and vitamins
and make hormones. Cholesterol and triglycerides avoid water, so they can’t travel through
blood themselves. This is why they combine with proteins to make lipoproteins that can move
throughout your body.

You’ll recognize some lipids by their nicknames: HDL (high-density lipoproteins) and LDL (low-
density lipoproteins).

Your liver and small intestine make HDL, which carries cholesterol into lipoproteins or your
liver. Your liver converts it to bile acid so you can get rid of it. HDL (the “good” cholesterol) also
fights inflammation, blood clots and oxidation.

Your liver makes VLDL (very low-density lipoproteins), which takes triglyceride from your liver
to other cells. When VLDLs drop off triglycerides and cholesterol, they get denser and become
LDL or the “bad” cholesterol. LDLs can stick to your artery walls and make it harder for blood
to get through your arteries.

FUNCTION

What do lipids do?

Various lipids have different functions. These include:

 Triglycerides transport and store energy.

 Steroid hormones send messages in cells.
 Bile salts from cholesterol help digest fat.
 Fatty acids metabolize to create energy.
 Phospholipids and cholesterol make biological membranes. Phospholipids are like
gatekeepers that let some molecules into your cells but require others to come in with
proteins.

ANATOMY

Where can I find lipids?

Lipids are in your blood and throughout your body. They’re in every cell you have.

What are lipids made of?

Oxygen, carbon and hydrogen bond to each other to form lipids. Lipids like cholesterol are part
of your cell membranes. They give your cells structure and allow substances to go in and out of
your cells.

CONDITIONS AND DISORDERS

What are the common conditions and disorders related to lipids?

Your body can produce the amount of lipids it needs. However, sometimes, it produces the
wrong amount of lipids. This can cause:

 Hyperlipidemia (high lipid levels).

 Hypolipidemia (low lipid levels).
 Hypercholesterolemia.
 Atherosclerosis.
 Coronary artery disease.
 Hypertriglyceridemia.

Common signs or symptoms of lipid conditions

Most people with high lipid levels don’t have symptoms. However, very high cholesterol can
show up as:

 Xanthelasma (bumps on the skin near your eyes).

 Xanthoma (cholesterol deposit under your skin).
 A white ring in your eye (corneal arcus).

What are common tests to check your lipid levels?

Your healthcare provider may order a lipid panel to measure the amount of cholesterol and
triglycerides in your blood. To get the most accurate results, you’ll need to avoid eating
anything (fast) for 12 hours before a provider takes a blood sample for the test. It’s easier to do
this early in the morning because you don’t eat while you sleep.

For accurate results, don’t take this test when you’re sick or stressed. Also, avoid taking
medicine that affects your lipid levels before the test. If you’re not sure whether any
medications or supplements you take can affect the results, ask your provider.

A lipid panel or lipid profile tells a provider if you’re at risk for cardiovascular (heart and blood
vessel) disease. Higher levels of total cholesterol (more than 200 milligrams per deciliter or
mg/dL) and LDL (more than 130 mg/dL) mean you have a higher risk of heart disease. People
with a history of coronary artery disease or stroke have more aggressive LDL targets because
they have a higher risk of further cardiovascular disease.

Higher levels of HDL (60 mg/dL) are good because HDLs remove LDLs from your body.

What are common treatments for lipids?

If you have high levels of lipids that put you at risk for heart disease, your provider may
prescribe cholesterol-lowering medication, such as:

 Statins.
 Bile acid sequestrants.
 Fibrates.
 PCSK9 inhibitors.

CARE

Simple lifestyle changes/tips for healthy lipid levels

To keep the bad lipids in your blood low or lower them if they’re too high, you can:

 Eat fewer calories.

 Eat less saturated fat, such as full-fat dairy foods.
 Choose whole grains, fruits, vegetables and reduced-fat dairy products.
 Exercise regularly.
 Sleep seven to nine hours every night.
 Strive for a weight that’s healthy for you.
 Stop using tobacco products. They can affect your HDL and LDL levels.

A note from Cleveland Clinic

Lipids have important functions in your body’s cells, but having too much of some lipids can
increase your risk of heart disease or cause other issues. Talk with your provider about your
risk of heart and blood vessel diseases. They can tell you how often you should get a lipid panel
to test your lipid levels. Also, they can help you get your numbers into a normal range.