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PROTEINS

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Main Ideas p121-140

Proteins are amino acids linked together by peptide bonds

Dehydration synthesis

Primary structure

Conformation of least energy

α-helix, β-sheet, helix-loop-helix e.g.

May have unstructured areas in between.

Secondary structure

Domains

Quaternary structure

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The central dogma of biology

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What are the structural levels of organization of proteins?

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a polypeptide chain

The linear sequence of AA’s in a protein constitutes the primary structure

of the protein

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Figure 4-2 Essential Cell Biology (© Garland Science 2010)

Remember: All 20 amino acids have a similar structure

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Aspartate

Isoleucine

How amino acids

interact with water

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Secondary

Structures

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tertiary structure

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(A) Cytochrome b562, a single-domain protein involved in electron transport in mitochondria. (B) the NAD-binding domain of the enzyme lactic dehydrogenase (C) the variable domain of an antibody

from the drawings of Jane Richardson

Tertiary structures are diverse

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Proteins made of more than one polypeptide subunit also have quaternary structure

Hemoglobin

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Figure 4-26 Essential Cell Biology (© Garland Science 2010)

The shape of extracellular proteins is often stabilized by disulfide bonds

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Four Levels of Protein Structure

    • Primary
    • Secondary
    • Tertiary
    • Quaternary

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Four Levels of Protein Structure

& bonds that contribute to the structures

    • Primary (peptide bonds)
    • Secondary
    • Tertiary
    • Quaternary

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Four Levels of Protein Structure

& bonds that contribute to the structures

    • Primary (peptide bonds)
    • Secondary (H bonds)
    • Tertiary
    • Quaternary

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Four Levels of Protein Structure

& bonds that contribute to the structures

    • Primary (peptide bonds)
    • Secondary (H bonds)
    • Tertiary (non-covalent & covalent (disulfide) bonds)
    • Quaternary

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Four Levels of Protein Structure

& bonds that contribute to the structures

    • Primary (peptide bonds)
    • Secondary (H bonds)
    • Tertiary (non-covalent & covalent (disulfide) bonds)
    • Quaternary (non-covalent & covalent bonds between subunits)

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What specifies the correct structure of a protein?

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What specifies the correct structure of a protein?

How do we know that a “correct” structure exists?

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Figure 4-5 Essential Cell Biology (© Garland Science 2010)

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This is reversible!

Denaturation by adding: 8M Urea to disrupt H2 bond and solvate hydrophobic side chains

β-Mercaptoethanol to disrupt disulfide bonds

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What can we conclude?

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Main Ideas p141-167

coenzyme and cofactor

Retinal

Heme

enzymatic pathway regulation

allosteric or conformational changes

prevent toxic buildup by regulating the beginning of a pathway

kinases, GTP binding proteins, protein complexes and proteolysis

Techniques

  • Lysis
  • Centrifugation
  • Chromatography
  • electrophoresis

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Molecular Chaperons

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Even a single change in the primary structure can alter the function of the protein

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Soluble, normal form

of a prion protein

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Insoluble, abnormal form

Soluble, normal form

of a prion protein

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Stanley Prusiner

Nobel Prize, 1997

Prion: proteinaceous infectious particle

Responsible for transmissable spongiform encephalopathies such as scrapie (sheep), kuru (cannibalistic tribe of New Guinea), Creutzfeld-Jakob disease (CJD), bovine spongiform encephalopathy (BSE, “mad cow disease”)

The abnormal form of the protein converts the normal protein to the abnormal form!

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  1. Frontal cortex ; spongiform degeneration

  • Plaques of Prion protein deposits.

Scale bar, 50 µm. Photomicrographs from S. DeArmond

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How would you regulate this pathway?

1 2 3 4 5

A B C D E F

6 7

G H

Enzyme 1 catalyzes the conversion of substrate A into Product B

Enzyme 2 catalyzes the conversion of substrate B into Product C

Enzyme 3 catalyzes the conversion of substrate C into Product D

Enzyme 4 catalyzes the conversion of substrate D into Product E

Enzyme 5 catalyzes the conversion of substrate E into Product F

Enzyme 6 catalyzes the conversion of substrate D into Product G

Enzyme 7 catalyzes the conversion of substrate G into Product H

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How would you regulate this pathway?

1 2 3 4 5

A B C D E F

6 7

G H

Product F inhibits enzyme 4

Product H inhibits enzyme 6

Product D inhibits enzyme 1