Biochemistry

545/645
September 29th, 2022
 Hemoglobin Mechanism – Structurally Distinct States of
Deoxy and Oxy Hemoglobin
DeoxyHb
T (Tense) State
Binding of O2 shifts the
relative orientation of
α2 α1β1 to α2β2 by 15°

α1

β1
β2

OxyHb R (Relaxed) State

15°
Hemoglobin Mechanism – Movements at the Atomic
Level
F helix
Leu
protoporphyrin
ring

Leu His iron

O
O

Leu a puckered iron to one

that’s flush

Val
Ionic Bonds Stabilizing Hemoglobin in the T State

F
heme H

proximal His H helix

F helix
position 87

F helix

F helix
 Hemoglobin Mechanism – More Movements at the
tissues Atomic Level lungs allosteric
regulators
“allo” ”steric”
“other” ”site”
O2 H +
O2

O2 H+
O2

Tense “T” Relaxed ”R”

pK~8 His 146 pK~6 His 146

deoxy hemoglobin oxy hemoglobin

Shift to oxyHb
associated with loss
of a proton and visa
versa
 Hemoglobin Mechanism – Allosteric Effectors
O lungs
2
(Protons & CO2 )
H+
lower pH, right shift
increased Kd, lower affinity
H+
O2

tissues

H+
~35%
Actively
~20%
respiring
H+

tissues are
producing H+
organic acids CO2 reacts with
and CO2 amino termini of
α globin chains
decreasing
affinity for O2
H2O + CO2 H2CO3 HCO3- + H+
 Hemoglobin Mechanism – Allosteric Effectors (2,3-
bisphosphoglycerate)
CO2
Binding of 2,3-
H – C – PO4 BPG stabilizes
deoxy state –
lowers affinity
CH2 – PO4 for O2
2,3 BPG – Net charge at pH 7: -5

Individuals
adapted to
lower pO2 at
higher HbF has a lower affinity for 2,3-
altitudes BPG and therefore a higher
have affinity for O2 than maternal HbA
elevated 2,3-
BPG
Hemoglobin Mechanism – Allosteric Effectors (Nitric
Oxide, NO)

Nitric oxide can bind to

free thiols in oxyHb –
allosteric switch to
deoxyHb releases NO
which can dilate blood
vessels.
 Hemoglobinopathies
Evolution of the Globin Gene Family
Ψ – nonfunctional pseudogenes

40%
17%
sequence
sequence
identity
identity

multicellular organism
The globin gene family is
thought to have evolved
through a series of gene
duplications and random
mutational drift
single celled
organism
Hemoglobinopathies – Population Studies on Hemoglobin
Variants

1 in 800 individuals have

variant forms of HB

~400 variants identified,

200 of which show no
clinical symptoms
 A 26-yo woman presents to the ER with lower extremity pain, pleuritic L chest
and upper quadrant pain, dry cough, and a temperature of 38.2°. Her lungs
were clear and an abdominal exam reveals generalized guarding but no
organomegaly. Chest X-ray is clear, and O2 saturation is 94%; WBC is
12,000 (12% bands); Hgb =8.5 gm (reticulocyte 15%). She’s admitted and
started on broad spectrum antibiotics/parenteral narcotics.
Norm. 37°
Norm. 11,300 Norm. Hgb 12.3-15.3 g/dl; 2.5% retics

Periodic/unpredictable episodes of pain due to

vaso-occlusive crisis

Serious infections

Chronic end organ damage

Strokes

Chronic anemia (hemolysis, splenic or hepatic

sequestration crises, aplastic)

Narcotic addiction
 Hemoglobinopathies – Sickle Cell Hemoglobin (HbS)
Incidence, World-Wide Distribution, and Malarial
Resistance
Incidence of sickle cell trait in
African American population ~8%
~ 40% in Equatorial Africa, Saudi
Arabia and India

HbS, a balanced
polymorphism with
good and bad
characteristics
Plasmodium falciparum
 Hemoglobinopathies – HbS, Structure/Function
Relationships
HbS, Val for Glu substitution
at position 6/7 of β globin
chains

Val-6

Tendency to polymerize is
more pronounced for
deoxyHb, favored by low pH
Cells infected with Plasmodium falciparum presumably
have lower pO2 and decreased pH which promote
sickling - ensuing events disrupt parasite’s life cycle
 Protein Structure/Function: Immunoglobulins
antigen
binding
site
heavy chain

hinge region
variable regions

light chain

Antibodies are proteins

constant regions • 4 polypeptide chains - 2 light
chains 2 heavy chains
• 4 chains held together by
disufide bonds
 Glossary
Ligand – any molecule that binds to a protein
Antigen – any molecule recognized and bound by an antibody
components of the adaptive immune system

Epitope – the specific part of an antigen recognized by an antibody

 Regions of Antibodies
Variable regions have
different amino acid
sequences from one hypervariable region
antibody to another

variable region

variable region
hypervariable
region
Constant regions have
“unvarying” amino
acid sequences from
one antibody to
another
 Antibody Fragments

cleavage
with
Fab Fab
papain
fragments fragments

Fc
fragment
B lymphocyte Development
 Antibody Trafficking
IgM IgG

Proteins are
Default pathway –
directed to
proteins in the
the secretory
lumen of the ER
pathway by a
are secreted from
signal at their . cells
amino .
terminus Proteins with
(signal signals that
peptide) anchor them in
the ER membrane
can end up at the
plasma
membrane
Secreted Proteins (IgG)