9/8/2016

Learning Objectives

Essential Biochemistry
Third Edition
Charlotte W. Pratt | Kathleen Cornely

Lecture Notes for

Chapter 5
Protein Function
09-09-16 (F)

To be able to understand structure and

function relationship
To be able to discuss how the quaternary
structure of hemoglobin contributes to
hemoglobins ability to transport oxygen
over long distances

Oxygen transport:
http://www.youtube.com/watch?v=5LjLFrm
KTSA&feature=related

Copyright 2014 John Wiley & Sons, Inc. All rights reserved.

Proteins serve a variety of functions.

Proteins serve a variety of functions.

Transport

Motor function

Myoglobin transports O2 throughout muscles.

Hemoglobin transports O2 in blood.

Structural
Actin forms microfilaments in cells.
Tubulin dimers constitute microtubules.
Keratin filaments constitute the bulk of animal
hair.
Collagen is a major protein in connective
tissue.
2014 John Wiley & Sons, Inc. All rights reserved.

Myosin interacts with actin to facilitate

muscular movement.
Kinesin moves along microtubules to support
a variety of cellular functions.

Other functions of proteins

Catalysis
Immunity
Regulation of gene expression
2014 John Wiley & Sons, Inc. All rights reserved.

9/8/2016

Why focus on
myoglobin and hemoglobin?

Myoglobin is a classical globular

protein.
Space-filling representation

Healthy red blood cell

Ribbon diagram with heme in purple

Sickled red blood cell

O2 transport is critical for sustaining life.

Hemoglobin mutation can possibly lead to disease.
Characteristics about O2 binding to myoglobin and
hemoglobin are observed in many areas of
biochemistry.
2014 John Wiley & Sons, Inc. All rights reserved.

Heme is a prosthetic group.

Prosthetic group =
organic molecule
bound to protein
that aids protein
function
Heme is a
porphyrin that
chelates iron for
oxygen transport.
2014 John Wiley & Sons, Inc. All rights reserved.

What is
heme?
2014 John Wiley & Sons, Inc. All rights reserved.

Myoglobin transports O2 via the

Fe in heme.

O2

His residues play a

key role in anchoring
both O2 and iron.
Anemia is often
treated with iron
supplements or an
iron-rich diet.

2014 John Wiley & Sons, Inc. All rights reserved.

9/8/2016

Myoglobin binds to O2 in a
hyperbolic trend.

O2 binding to the heme group of

myoglobin exhibit a hyperbolic curve.

Fractional Saturation (Y):

the proportion of myoglobin
molecules that have bound O2

Y=

Bound Mb
Total Mb

Y=

[MbO2]
[Mb] + [MbO2]

Hyperbolic data is common in biochemistry!

2014 John Wiley & Sons, Inc. All rights reserved.

2014 John Wiley & Sons, Inc. All rights reserved.

Remember!

Mb and Hb are only ~18% identical in

primary sequence.

Proteins have four possible levels of

structure.
Primary sequence
Invariant

Identical in all

Identical in Hb

Secondary: alpha helices and beta sheets,

coils and loops

Tertiary: 3D fold
Quaternary: interaction of multiple subunits
2014 John Wiley & Sons, Inc. All rights reserved.

2014 John Wiley & Sons, Inc. All rights reserved.

9/8/2016

Mb and Hb are similar in their

secondary and tertiary structures.
Heme

Even though
myoglobin and hemoglobin have
only ~18% identical residues,
their secondary and tertiary
structures overlap almost
perfectly when superimposed!

Myoglobin
-Subunit of Hemoglobin
-Subunit of Hemoglobin

The similarities in structure and

sequence between myoglobin and
hemoglobin indicate a common
evolutionary origin.

Hb has quaternary
structure, but Mb
does not.

2014 John Wiley & Sons, Inc. All rights reserved.

KEY CONCEPTS: Section 5-1

Myoglobin binds to O2 in a hyperbolic
trend.
The similarities in structure and sequence
between myoglobin and hemoglobin
indicate a common evolutionary origin.

2014 John Wiley & Sons, Inc. All rights reserved.

Oxygen binds cooperatively to Hb.

Dotted line represents O2

binding to myoglobin (hyperbola).
Solid line represents O2
binding to hemoglobin (sigmoid).

Note:
Sigmoidal data are
indicative of cooperativity.

Cooperativity:
Binding of O2 to one subunit induces easier binding to other subunits.
2014 John Wiley & Sons, Inc. All rights reserved.

2014 John Wiley & Sons, Inc. All rights reserved.

9/8/2016

Quaternary
structure
defined as
heterotypic:
with different
subunits

Why hemoglobin and

not myoglobin
performs the role of
transporting oxygen
over long distances?

Fe2+
(ferrous);
reduced form

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Differences in the Binding Affinity of two

O2 Binding Proteins

Sigmoidal curve exhibits

cooperative binding. It
exhibits allosteric
interactions.

Hyperbolic
curve

Hemoglobin exhibits
cooperative binding,

The binding of one molecule of

O2 to hemoglobin causes a
conformational change to
hemoglobin which enhances the
ability of hemoglobin to bind
more O2 molecules. This property
of hemoglobin is known as
"cooperative binding."

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Binding and Release of Oxygen Induces

Conformational Change

Hemoglobin exhibits
cooperative binding

O2

O2

O2
O2

Differences in the Binding Affinity of two

O2 Binding Proteins

Biological
significance ?

The Ability of Hemoglobin to Transport

Oxygen Over Long Distance
At 15 kPa
hemoglobin is
95% saturated
with oxygen

At 5 kPa
hemoglobin
is 75%
saturated
with oxygen
argosymedical.com

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What structural difference

accounts for the ability of
hemoglobin to transport oxygen
over long distances compared to
myoglobin?

Bohr Effect and O2 Transport

What is happening biochemically when you breathe?

From
Metabolism

+ H2O

2014 John Wiley & Sons, Inc. All rights reserved.

What is the effect of 2,3BPG to oxygen binding?

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KEY CONCEPTS: Section 5-1

O2 can bind cooperatively to hemoglobin
as the protein shifts from the deoxy to the
oxy conformation.
The Bohr effect and BPG modulate
hemoglobin function in vivo.

2014 John Wiley & Sons, Inc. All rights reserved.