x ray crystallography

34 g/dL = Hb in RBC
Approximately 1.34 mL oxygen is bound by each
gram of Hb.
64,000 D
O2 transport, NO transport to modulate vascular
dilation, CO2 transport from tissues to lungs
Heme ferroprotoporphyrin IX
Double bonds red
4 heme groups (each can combine with 1 molecule of
O2) = 4 molecules O2

Globin 2 pairs / 4 globin chains (141-146 aa)

Globin

4 globin chains
8 helices A to H
7 non helical segments / sequences
NA, AB, BC, CD, DE, EF, FG, GH, HC
Heirarchy of protein structure
A1B2 / A2B1 = between dimers
A1B1 = front
A2B2 = back

 Heme = suspended between E and F

Ferrous = proximal his F8 and distal his E7
(swings in and out for O2)
Hydrophilic vs. hyrdphobic amino acids
Each globin chain has a heme group attached.
Each heme molecule can carry one molecule
of oxygen.
F cells = w/ HbF in adult

HbA1c
Glycation is a posttranslational modification
formed by nonenzymatic binding of various
sugars with globin chain amino groups.
The most glycated Hb is HbA1c, in which
glucose attaches to the N-terminal valine of
the B-chain.

Hemoglobin Synthesis
Heme & Globin Biosynthesis
Mature RBC can no longer make Hb.

Hb assembly
After their release from ribosomes, each
globin chain binds to a heme molecule and
they pair off.

 Bohr effect shift in the curve due to pH

Shift to the left

Lowered body temperature due to external
causes
Multiple transfusions of stored blood with
depleted 2,3 DPG
Alkalosis
Presence of metHb, carboxyHb or other Hb
variants

Shift to the right

High fever
Acidosis
Hypoxia high altitude, pulmonary
insufficiency, CHF, severe anemia, and cardiac
right to left shunt

Myoglobin
Rhabdomyolysis

Hemoglobin

Birth = 15 20 g/dL
2 months = 9 14 g/dL
10 years old = 12 15 g/dL
Adult = 12 16 (F); 13 18 (M)
After 50 years of age = slight decrease
Higher in the morning and lower in the evening
Lower if lying down
Increased in smokers; high altitude

Forms of Hb
oxyHb
deoxyHb
carbaminoHb

Abnormal Hb pigments
carboxyHb = Hb + CO; reversible
metHb = Fe++ Fe+++; reversible
sulfHb = irreversible due to sulfonamides,
aromatic amines
carboxysulfHb

Fe metabolism

 70% = Fe ++ = heme
5 % = Fe++ + Mb O2 transport in muscles
Hb (tetrameric); Mb (monomeric)

25% = ferritin; hemosiderin

<5% = heme bound Fe for cyp450 oxidase
Fe++<--Fe+++

Dietary Fe (minimally bioavailable)

10-20 mg/day absorbed: 1-2 mg/day
Heme Fe (meat) absorbed more efficiently
Non heme (non meat)
Enhance absorption on non heme Fe:
ascorbate, citrate, organic / amino acids

Interfere with absorption of non heme Fe:

phytates, polyphenols, phosphates, oxalates, calcium

Cereals fortified with Fe

Fe absorption (from food)

Duodenum & upper jejunum
ferric
DCYTB
non heme Fe soluble form
duodenum specific cytochrome b-like protein
Uptake of Fe occurs on heme carrier protein1
in apical membrane of duodenal enterocyte.
Heme Fe + enterocyte binding


heme oxygenase

heme Fe++, CO, bilirubin IX-a

degrade
Transport:
Fe + apical DMT1 duodenal epithelium
Divalent metal transporter 1
ferroportin (basolateral transport protein)
Fe++ basolateral portal circulation

 Ferroportin works with hepahestin. (Cucontaining Fe oxidase)

hepahestin
Fe ++ Fe+++ to transferrin

reoxidation
Enterocyte (stool) stored ferritin
Hepcidin (antimicrobial from liver) is a
negative regulator of intestinal Fe absorption
and release from macrophages.
Hepcidin + ferrroportin receptor
degradation of ferroportin & Fe trap in
intestine.

Increase in hepcidin synthesis

If transferrin saturation is >
50 % in females or 60 % in
males.

Decrease in hepcidin synthesis

If Fe saturation is low.

Transferrin
Transports Fe+++ to hematopoietic and other
tissues with transferrin receptors (in
normoblasts or rapidly dividing cells).
Endocytosis endosome with Fe loaded
transferrin
Fe release from transferrin by acidification of
endosome to pH 5.5

 Fe transport across endosomal membrane by

DMT1 and used in synthesis of Fe-containing
protein.
Excess Fe stored as ferritin and hemosiderin.
Fe excretion ineffective. Absorption is
therefore regulated.
Amount of Fe absorbed is inversely
proportional to Fe stores & rate of
erythropoiesis.

Fe loss
Mainly be feces (1 mg/day)
Minimal loss perspiration, skin/dermal
exfoliation
Lactation and menstruation 1 mg/day

Fe cycle and transport

Fe absorbed GI
Fe transport to bone marrow inserted to
protoporphyrin IX
Fe in RBC circulates (Fe++)
Fe from senescent RBCs macrophage
reuse

Ferrokinetics
Transferrin, transferrin receptors, ferritin =
regulated by IRP (iron-responsive protein)
Plasma transferrin (hepatocytes) Fe
transport from enterocyte of duodenum
transferrin receptors on normoblasts
Transferrin half-life of 8 days; beta fraction in
electrophoresis
Each NH2 & -COO terminus bind to Fe+++

 Bicarbonate locks Fe in place with in

transferrin (bridgin ligand)
Transferrin gene long arm of chromosome 3
Transferrin exists as:
apotransferrin (single chain of clycoprotein
without Fe)
Monoferric
Diferric form

Transferrin receptor

Long arm of chromosome 3

Glycoprotein dimer
On all cells except mature RBCs
Provides transferrin-bound Fe across into
normoblasts
Release of Fe from transferrin
Increased in normoblasts, placenta, liver
Can bind 2 molecules of transferrin

Transferrin receptor
Increase transport of Fe & pH 7.4 = transferrin
receptor selectively binds diferric > monoferric
transferrin or apotransferrin.
Control of transferrin receptor biosynthesis:
major mechanism for regulation of Fe
metabolism; synthesis induced by Fe
deficiency

 If transferrin is fully saturated Fe deposited

in liver.
If congenitally absent, Fe is absorbed by
intestine & accumulates in liver, pancreas,
spleen microcytic hypochromic anemia.
IRP binds to IRE (iron responsive elements)
when Fe supply decreases.

Fe storage
Ferritin accessible
Hemosiderin partially degraded or
precipitated form of ferritin
Apoferritin protein component of ferritin
w/o Fe; with 24 L (light) and H (heavy)
subunits.
L subunits liver and spleen (chromosome 19)
H subunits heart (no ferritin; chromsome 11)

Hemosiderin degradation product of ferritin

Fe storage
Increased Fe stores increased hemosiderin:
ferritin
Ferritin and hemosiderin stores : Liver > bone
marrow, spleen

Lab Assessment of Fe metab

(p.131, table 11-2)
Serum Fe measure of ferric bound to serum
transferrin (does not include free serum Hb
Fe)
Chromogen spectro
Increased in am ; low in pm
Decreased in IDA, inflammation, acute infection,
certain immunizations, post MI

Lab assessment
UIBC unsaturated Fe binding capacity
TIBC indirectly - chemical means; directly
transferrin
% transferrin saturation = serum Fe/TIBC x 100
Plasma ferritin low in early IDA; high acute
phase reactant

sTR (serum transferrin receptor)

Immunoassay
high if cells lack Fe ; ineffective erythropoiesis
Low in chronic diseases
sTR: log serum ferritin (sTR-F index)
IDA vs. anemia of chronic inflammation

RBC protoporphyrin
Increased heme production (IDA; Fe use is
blocked)
Combines with Zn when Fe is unavailable
fluorescent Zn protoporphyrin
High = disorders of heme synthesis, IDA, Pb
poisoning, sideroblastic anemias, chronic
diseases

Tissue Fe

BM / liver biopsy (Prussian blue)

Sideroblasts nRBC
Siderosomes granules
Siderocytes reticulocytes in BM with Fe
not normal; Fe granules in retics are not seen in
light microscopy.