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    Protein Folding

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    9374 DIVYATA PATIL
    This document discusses protein denaturation and renaturation. It defines denaturation as the process where a protein loses its native 3D structure and discusses common denaturants like heat, acids, bases, and detergents. It also summarizes Anfinsen's classic experiment which showed that a protein's primary structure contains all the information needed to refold it into its correct 3D structure after denaturation.

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    Protein Folding

    Uploaded by

    9374 DIVYATA PATIL
    25 views32 pages
    This document discusses protein denaturation and renaturation. It defines denaturation as the process where a protein loses its native 3D structure and discusses common denaturants like heat, acids, bases, and detergents. It also summarizes Anfinsen's classic experiment which showed that a protein's primary structure contains all the information needed to refold it into its correct 3D structure after denaturation.

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    Protein folding

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    This document discusses protein denaturation and renaturation. It defines denaturation as the process where a protein loses its native 3D structure and discusses common denaturants like heat, acids, bases, and detergents. It also summarizes Anfinsen's classic experiment which showed that a protein's primary structure contains all the information needed to refold it into its correct 3D structure after denaturation.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    25 views32 pages

    Protein Folding

    Uploaded by

    9374 DIVYATA PATIL
    This document discusses protein denaturation and renaturation. It defines denaturation as the process where a protein loses its native 3D structure and discusses common denaturants like heat, acids, bases, and detergents. It also summarizes Anfinsen's classic experiment which showed that a protein's primary structure contains all the information needed to refold it into its correct 3D structure after denaturation.

    Copyright:

    © All Rights Reserved
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    Download as pdf or txt
    of 32

    • Denaturation and Renaturation of proteins;

    • Denaturants and their mode of action;


    • Anfinsen’s classical experiment;
    AMINO ACID
    • Denaturation is the process in which protein loses its
    native confirmation
    • It basically involves the disruption and possible destruction
    of both the secondary and tertiary structures
    • It mainly breaks the covalent and non covalent bonds, but
    they are not strong enough to break peptide bonds
    • So the primary structure remains intact
    • Precipitation or coagulation or flocculation of protein
    occurs
    • Heat
    • Strong acids
    • Strong bases
    • Detergents
    • Reducing agents
    • Heavy metal ions
    • Organic solvents
    • Solutes
    • Most proteins can be denatured by heat, which
    affects the weak interactions of the proteins in a
    complex manner
    • Interactions affected: Hydrogen bond
    Nonpolar (Hydrophobic) bond
    • As Kinetic energy increases, molecules vibrate
    rapidly and violently
    • Protonation and Deprotonation of the side groups
    of protein occurs
    • Interactions affected: Salt bridges,
    Hydrogen bond,
    Electrostatic interaction.
    • Detergents are amphipathic molecules
    • Interactions affected: Hydrogen bonds,
    Non-Polar bonds (Hydrophobic)
    • Disulphide bonds are formed by oxidation of
    sulfhydryl groups on cysteine
    • Reducing agent reduces the disulphide bonds to
    sulfhydryl groups & breaks intra and interchain
    disulphide bonds
    • Interactions affected: Non-covalent bonds
    (Disulphide bonds)
    Reducing agent:
    Beta- Mercaptoethanol
    Urea

    Reductants add Hydrogen atoms to make the thiol


    group (-SH)
    • Heavy metal usually contains: Hg+2, Pb+2 & other
    metals with high atomic weights.
    • Heavy metal salts denature proteins the way acids
    and bases denature.
    • Interactions affected: Non-covalent bonds
    (Disulphide bonds)
    The reaction of a heavy metal salt with a protein
    usually leads to an insoluble metal salt protein
    • They act primarily by disrupting the hydrophobic
    interactions that make up the stable core of
    globular proteins
    • New hydrogen bonds are formed instead
    between new alcohol molecule and protein side
    chains
    • Interactions affected: Hydrogen bonds,
    Polar (Hydrophillic bonds)
    • Reversible denaturation is
    called Renaturation.
    • Example: Hemoglobin
    undergoes denaturation in the
    presence of salicylate. By
    removal of salicylate,
    hemoglobin is renatured
    • Example: Anfinsen’s
    Experiment
    • The classic experiment, carried out by Christian
    Anfinsen in the 1950s, provided the first evidence
    that the amino acid sequence of a polypeptide
    chain contains all the information required to fold
    the chain into its native, three-dimensional
    structure.
    • Later experiments proved that the primary
    structures determines the conformation of the
    protein
    • In this following experiment he used a particular
    type of enzyme called Ribonuclease
    • Urea: Disrupts the non-covalent bonds such as
    ionic interactions and hydrogen bonds
    • Mercaptoethanol: Breaks down the disulphide
    bonds by an oxidation- reduction reaction
    • He conducted 3 experiments.
    • When the native enzyme was treated with excess
    Beta-Mercaptoethanol and 8.0M Urea, all disulfide
    bonds and hydrogen bonds were broken and the
    protein denatured. When the denaturing agents
    were removed the enzyme eventually reformed its
    original tertiary structure
    • When Beta-Mercaptoethanol was removed first, an
    inactive enzyme was formed. This is because the
    improper disulfide bonds were formed
    • When trace Beta-
    Mercaptoethanol was added
    to the scrambled enzyme,
    the correct tertiary structure
    was eventually reformed.

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