Science of Living System

Nihar Ranjan Jana

School of Bio Science
Email: nihar@iitkgp.ac.in
Tel: 03222-260802
 Central Dogma of Life
DNA: Storage Medium
Polymer of nucleotides

CCTGAGCCAACTATTGATGAA

RNA: Transmission Medium

Polymer of nucleotides

CCUGAGCCAACUAUUGAUGAA

Protein: Molecular Machines

Polymer of amino acids

PEPTIDE
 What is a Protein
Hierarchy of Protein Structure
Proteins are polymers built up from
20 different amino-acids linked by
peptide bonds

Ala,Cys,Asp,Glu,Phe,Gly,His,Ile,Lys,Leu
A C D E F G H I K L
Met,Asn,Pro,Gln,Arg,Ser,Thr,Val,Trp,Tyr
M N P Q R S T V W Y

The 3D-structure of a protein is

encoded in its amino acid sequence
(primary structure)!
 Primary Structure of Proteins
The primary structure of a protein is its amino acid sequence
Amino Acid

Amino acids have a common chemical

structure - A tetrahedral sp3 carbon (Cα)
with four different functional groups:

1.Amino group
2.Carboxyl group
3.H-atom
4.Side chain (R) with distinct chemical
property

There are 20 common amino acids. The

R group (also known as side chain),
attached to the α carbon is different in
each amino acid
 All Amino Acids in Protein Have the “L-form”

H-atom is coming out of the whiteboard. Looking down the H-Cα bond
from the H-atom, the L-form amino acid has CO, R and N going in a
clockwise direction. The L-form reads “CORN” in clockwise direction.
The 20 Common Amino Acids of Proteins
 Amino Acid with Special Characteristics
cysteine

cysteine cystine

Reversible formation of a disulfide bond by the oxidation of two

molecules of cysteine. Disulfide bonds between Cys residues
stabilize the structures of many proteins
Formation of the Peptide Bond

1 2 3 4 5 6 Polymer: Polypeptide
and Protein
Monomers Peptide bonds
(amino acids)
Formation of the Peptide Bond
The amide plane: partial double bond
character of the peptide bond
 Cα Cα

Cα Cα
TRANS

H H
Cα Cα
CIS

Cα Cα
Torsion angles:
Φ (phi) and Ψ (psi)

The N—Cα and Cα—C bonds can rotate, and designated as Φ and Ψ angles
respectively. The peptide C—N bond is not free to rotate. Other single bonds in
the backbone may also be rotationally hindered, depending on the size and
charge of the R groups
φ and ψ Torsion Angles are the Only Degrees
of Freedom for the Backbone
 Ramachandran Plot: The φ - ψ Space
Φ Ψ

ω
G. N. Ramachandran

+180

Ψ (psi)

-180
-180 Φ (phi) +180
 Ramachandran Plot
α-Helix

β-Strand
Ψ (psi)

Φ (phi)
Glycine Residues can Adopt Many Different
Conformations

• Glycine with only a H-atom as side chain can

adopt a much wider range of Φ-Ψ conformations
than the other residues

• It thus plays a structurally important role; it allows

unusual main chain conformations in proteins

• This is the main reasons why a high proportion of

Glycine residues are conserved among
homologous protein sequences
 Central Dogma of Life
DNA: Storage Medium
Polymer of nucleotides

CCTGAGCCAACTATTGATGAA

RNA: Transmission Medium

Polymer of nucleotides

CCUGAGCCAACUAUUGAUGAA

Protein: Molecular Machines

Polymer of amino acids

PEPTIDE
Proteins come in various shapes and sizes
The protein folding problem
- Consider a small protein with 100 residues.

- Cyrus Levinthal calculated that, if each residue can assume

three different conformations, the total number of structures
would be 3100, which is equal to 5 × 1047. If it takes 10-13 s to
convert one structure into another, the total search time would
be 5 × 1047 × 10-13 s, which is equal to 5 × 1034 s, or 1027 years
i.e. longer than the age of the universe!

- Clearly, it would take much too long for even a small protein to
fold properly by randomly trying out all possible conformations.

- The enormous difference between calculated and actual folding

times is called Levinthal's paradox.
 The 3D structure of a protein is encoded in its
primary sequence: Anfinsen’s Experiment

Thermodynamic hypothesis of
Protein Folding: The interactions
between the atoms in a protein control
the folding of the protein molecule into
a well-defined three-dimensional
structure.

In other words, the protein sequence

contains enough information required
for the proper folding of the protein
into its functional three-dimensional
structure.
 Anfinsen’s Experiment

If we understand HOW PROTEINS FOLD, we can predict their structure from

sequence! Then we can design proteins with novel functions.
Anfinsen’s Experiment
 Forces that stabilize a protein structure

Most important feature: The interior of proteins is hydrophobic!

The main driving force for folding water soluble globular protein
molecules is to pack hydrophobic side chains into the interior of
the molecule, thus creating a HYDROPHOBIC CORE and a
HYDROPHILLIC SURFACE.

Problem: How to create such a hydrophobic core from a

linear protein chain ???
Hydrophobic core formation drives
protein folding
The Protein Folding Game - Foldit
https://fold.it/portal/
Motifs of Protein
Structure
Protein Molecules are Organized in a Structural
Hierarchy

Primary

Secondary

Tertiary

Quaternary
 Secondary Protein Structure
Characterized by main chain NH and CO groups participating in H-bonds

Beta Sheet

Alpha helix
 Alpha Helix

Every 3.6 residues make one turn

The distance (pitch of helix)

between two turns is 5.4 Å

The C=O of residue ‘n’ is hydrogen

bonded to N-H of residue ‘n+4’

n n+1 n+2 n+3 n+4 n+5

Some Amino Acids are Preferred in α-Helices

Good helix formers:

Ala , Glu, Leu , Met

Less Preferred:
Pro, Gly, Tyr, Ser
 Alpha Helix: Right-handed or Left-handed?

Alpha helix can be – Right-

handed or Left handed

BUT, left handed helix is not

possible for L-amino acids
due to close approach of
the side chains and CO
group.

Right handed – most

commonly observed in
proteins.
 Helical Wheel Plot

C
 Hydrophobic

Hydrophilic

Charged

Totally buried Partially buried

Helical Wheel: Each

residue can be
Exposed
plotted every
360/3.6=100° around
a circle or spiral
 β-sheet
(Number of β-Strands are Involved)

β-sheet from several regions of the

chain; Each β-strand, typically 5-10 α-helix: from one
residues long continuous region
H-bonds are perpendicular to strands
Parallel and Antiparallel β-sheet

β-pleated sheet: ‘pleated’ because

side chains point up and down
alternatively
Mixed β-sheet
Hair Keratin
Hair Keratin
Chemistry of Straight and Curl Hair
 Polypeptide Chains Fold into Several Domains
•Fundamental unit of tertiary structure – DOMAIN
•Domain: polypeptide chain or a part of polypeptide chain that can
independently fold into a stable tertiary structure
•Domains are also units of function

Tertiary structure refers to the spatial arrangement of amino acid residues

that are far apart in the sequence and to the pattern of disulfide bonds.
 Quaternary Structure
Proteins containing more than one polypeptide chain exhibit a fourth level of
structural organization. Each polypeptide chain in such a protein is called a
subunit. Quaternary structure refers to the spatial arrangement of subunits
and the nature of their interactions.

The simplest quaternary structure is a dimer, consisting of two identical

subunits.
Quaternary Structure (higher order)
The α2β2 tetramer of human
haemoglobin. The structure of the two
identical α subunits (red) is similar to
but not identical with that of the two β
subunits (yellow).

Complex Quaternary Structure. The

coat of rhinovirus comprises 60 copies
of each subunits
 Solving Protein Structures
Only 2 kinds of techniques allow one to get atomic resolution
pictures of macromolecules

• Structure Function
• Structure Mechanism
• Structure Origins/Evolution
• Structure-based Drug Design
• Solving the Protein Folding Problem

H: α-helix
E: β-strand
QHTAWCLTSEQHTAAVIWDCETPGKQNGAYQEDCA
HHHHHHCCEEEEEEEEEEECCHHHHHHHCCCCCCC C: unstructured
 Importance of Protein Structure

Hemoglobin A: Val-His-Leu-Thr-Pro-Glu-Glu-Lys-
Hemoglobin S: Val-His-Leu-Thr-Pro-Val-Glu-Lys-

“sticky patch” causes hemoglobin S to agglutinate (stick together) and

form fibers which deform the red blood cell
 https://www.youtube.com/watch?v=qBRFI
McxZNM

Books Followed:

Biochemistry (Lubert Stryer)

Principles of Biochemistry (Nelson and Cox)