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Quaternary Structure

Quaternary structure refers to the arrangement of multiple polypeptide chains in a protein complex. Proteins can assemble into complexes through the association of identical or different subunits. Quaternary structure provides stability and allows the formation of large complexes through subunit interactions. Changes in quaternary structure, such as domain swapping, can lead to the formation of pathogenic protein aggregates associated with diseases like Alzheimer's. Understanding protein-protein interactions is important for controlling the assembly of protein complexes and preventing unintended aggregation.

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123 views

Quaternary Structure

Quaternary structure refers to the arrangement of multiple polypeptide chains in a protein complex. Proteins can assemble into complexes through the association of identical or different subunits. Quaternary structure provides stability and allows the formation of large complexes through subunit interactions. Changes in quaternary structure, such as domain swapping, can lead to the formation of pathogenic protein aggregates associated with diseases like Alzheimer's. Understanding protein-protein interactions is important for controlling the assembly of protein complexes and preventing unintended aggregation.

Uploaded by

Chiku Mtegha
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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Quaternary structure

• Assembly of multiple polypeptide chains


in one integral structure
• The arrangement of the subunits gives
rise to a stable structure
• Subunits may be identical or different
• A common shorthand for describing such
proteins is to use Greek letters for each
type of subunit, and subscript numeral to
specify numbers of units.
– A protein designated α2βγ consists of two
α units and one each of β and γ

1 = monomer 7 = heptamer
2 = dimer 8 = octamer
3 = trimer 9 = nonamer
4 = tetramer 10 = decamer
5 = pentamer 11 = undecamer
6 = hexamer 12 = dodecamer Petsko and Ringe
Escher
• Quaternary structure adds stability by decreasing the
surface/volume ratio of smaller subunit
• Simplifies the construction of large complexes
– viral capsids are often composed of multiples of 60
proteins
– 20s subunit of proteosomes contain four heptameric
rings (4 x 7 = 28 subunits)

Hepatitis B virus

proteosome degrades unfolded protein—cellular garbage disposal


Why are some enzymes so large
Small enzymes, e.g. hydrolases, may contain ~ 125 amino acids
Other enzymes, e.g. large dehydrogenase, may contain > 60,000 amino
acids

• Large structure provides rigidity necessary to orient the substrate and


key amino acids to enable catalysis
– e.g. extremely small proteins require metals or disulfides for stability
• Large surface area can funnel small substrates
to the active site—e.g. electric field gradient
• Substrates need to be shielded from the solvent
• Surface/volume ratio decreases with size, and
reduces the tendency to aggregate

Klapper et al, Proteins 1, 47 (1986)


Viral capsid
Virus comprises a genome made of either DNA or RNA, a protein shell
(“capsid”) around it, and lipid bilayer outside in some cases

A nucleic acid cannot code for a single protein molecule large enough to
enclose it. Therefore, many copies of short polypeptides must assemble
to build the capsid

Structural information of viral capsid can


lead to the rational design of antiviral drugs
– prevent viral infection by blocking the
interaction between viral protein and
cellular target
– prevent viral assembly by disrupting
quaternary association of viral capsid
proteins

Branden & Tooze, Ch 16


Arrangement of subunits
Most protein multimers have significant rotational symmetry in the
placement of the subunits

Potassium channel
Chemistry Nobel prize, 2003

Human immunodeficiency
virus aspartyl protease
Advantages of building protein complexes
• Easier to evolve—shorter genes
• Easier to transcribe and translate—quicker response
• Robust against error in transcription/translation (1 in 2000 amino acids)

• Additional layer of regulation


– Allosteric properties not present in monomer
– Many multiprotein complexes regulate their physiological function through
conformational changes
– Changes in quaternary structure can occur through conformational changes
within individual subunits or through reorientation of the subunits relative to
each other.
Hemoglobin v. myoglobin
Hemoglobin: heterodimer of dimer (α2β2)
Oxygen binding in hemoglobin is highly cooperative
cooperativity is achieved through domain rotation
compare with myoglobin, which is a monomeric protein

Eaton et al, NSB 6, 351 (1999)


Formation of quaternary structure
• The subunits are held together by both hydrophobic interactions and
ionic interactions between polar/charged amino side chains
– a quaternary structure may fall apart in high salt environment
• A monomer typically buries 600 – 5000 Å of surface
• Understanding protein-protein interaction is key to understanding and
controlling the formation of protein complexes

interface
residue
“Lock and key” at the dimer interface
Glutathione S transferases (GST) are dimeric enzymes that
neutralize nonpolar exogenous toxic compounds by
nucleophilic addition of glutathione

A dimeric quaternary structure is essential for function and


the active site is formed by amino acids from both subunits

The association relies on hydrophobic interaction in which


an aromatic ‘key’ from one domain inserts into a
hydrophobic pocket (‘lock’) of the other domain

Wongsantichon & Ketterman, Biochem J 394, 135 (2006)


E. coli RNA polymerase dimer
The N terminal domain (NTD) is thought to be involved in dimer formation

The interface consists of both polar (E32, T38, S50, and Q227) and
hydrophobic (F35, F8, L31, L39, I46) residues, which together form a
cluster that provides more stability than a single pair of polar–polar or
hydrophobic–hydrophobic interaction

Kannan et al, Protein Sci 10, 46 (2001)


3D domain swapping
• A common mechanism for forming oligomeric proteins from monomers
• Creates an interface between different polypeptide chains which is
identical to that seen within the monomer
• The interface for dimerization has already been evolved and optimized
• Monomers may exchange a secondary structural element (strand or
helix) or an entire domain

Bennett et al. Structure 14, 811 (2006)


Altering quaternary structure
• Engineer monomeric staphylococcal nuclease ultracentrifugation
into a dimer MW: 16 kDa x 2 = 32 kDa
• Deletion creates steric clash involving side
chains and promotes dimerization

Green et al, NSB 2, 746 (1995)


The details of the interface are maintained in the mutant
Protein G

Highly stable and compact proteins (e.g. immunoglobulin binding domain


of protein G) can change its topology and quaternary structure by a
few changes in core residues

Byeon, et al JMB 333, (2003)


p13suc1 Strand exchange
Natural proteins often dimerize by exchanging a
strand or a helix

hinge loop Domain-swapped structures are biologically


significant

In p13suc1, the hinge loop works as a loaded


molecular spring

Mutation in the hinge loop shifts the


monomer-dimer equilibrium

The binding of a ligand (e.g. phosphate and


phosphopeptide) also shifts the equilibrium in
wild type by altering the dynamic properties of
the native state ensemble

Schymkowitz et al, NSB 8, 888 (2001)


Domain swapped protein fibrils
may be involved in diseases
prion, hereditary hemorrhagic stroke disorder

prion disease mutations map to domain swapping helix

consecutive domain
swapping is also
possible leading to fibril
formation

prion dimer

Knaus et al, NSB 8, 770 (2001) Bennett et al. Structure 14, 811 (2006)
Deposition diseases
Deposition diseases are “conformation” diseases characterized by
aggregation of native proteins
–Amyloidic : ordered fibril like deposits of proteins
»Alzheimer’s, Parkinson’s, Huntington, Type II diabetes
»beta strands running perpendicular to the fibril axis
–Non-amyloidic fibrils or aggregates
»sick cell anemia
»serpinopathies

The proteins implicated in these diseases usually perform well-


characterized, essential biological functions under normal circumstances …
until things fall apart

A change in environment or genetic predisposition exposes a protein for the


growth of aggregates—unintended quaternary structure
Mechanism of fibril formation
3D domain swapping
End-to-end stacking
Cross-beta spine

Amyloids
Ordered fibrillar aggregations
Adopt a cross-β structure
Fiber diffraction, EM, etc.
1.8 Ǻ structure of yeast prion protein
Involve conformational changes
(Partial) denaturation
Proteolysis
All proteins may be induced
to form amyloid fibrils

Nelson et al, Nature 435,773 (2005)


Etiology of amyloid disease
Amyoid diseases are folding diseases—a change in the primary and/or
tertiary structure results in the formation of undesirable quaternary
structure

This is an example of the breakdown in the principle of “negative design”


Negative design : structural elements that disfavor all non-native structures,
including unfolded structure, oligomerization, alternate topologies

Q: How does a protein protect itself from


forming unintended aggregates?

We need understand the sequence-structure


relationship better to predict what sequences
are amyloidic

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