UNIVERSITÄTSMEDIZIN

NEUMARKT A. M. https://edu.umch.de
www.umfst.ro

CAMPUS HAMBURG PRESENTER: 2022 SEPTEMBER, 29th

Lecturer Zsolt KOVACS
Lecture 2. HETEROPROTEINS.
NUCLEOSIDE TRIPHOSPHATES
Contents
1. Aminoacids
1.1. Clasification of aminoacids
1.2. The biological role of aminoacids
1.3. Physical properties of aminoacids
1.4. Non proteinogenic amino acids
2. Peptide
2.1. Peptide of biological importance
3. Proteins
3.1. Proteins structure
3.1.1. Primary structure
3.1.2. Secondary structure
3.1.3. Tertiary structure
3.1.4. Quaternary structure
3.2. Clasification of proteins
3.2.1. Simple proteins
3.2.2. Heteroproteins
3.2.2.1. Glycoproteins
3.2.2.2. Lipoproteins
3.2.2.3. Chromoproteins
3.2.2.4. Nucleoproteins
Chromoproteins
1. Hemoglobin vs Myoglobin – General considerations
2. Heme structure
3. Oxygen binding to Myoglobin and Hemoglobin
3.1. Factors that influence oxygen binding to hemoglobin
3.1.1 pO2
3.1.2. Cooperative binding of oxygen to Hb - physiological significance
3.1.3. 2,3-Bisphosphoglycerate
3.1.4. Hydrogen Ions Promote the Release of Oxygen: The Bohr Effect
4. Carbon monoxide binding to Hemoglobin
5. Carbon dioxide transport
6. Pathological aspects
Nucleoproteins
1. General considerations
2. Pentose
3. Nitrogenous bases
4. Nucleosides
5. Nucleotides
6. Nucleic acids
6.1. DNA
6.2. RNA
Nucleoside triphosphates. Biochemical energy concepts

2
3.2.2. HETEROPROTEINS
3.2.2.1 Glycoproteins= carbohydrate group can be covalently attached to a protein to form a glycoprotein.
• 50% of the proteome consists of glycoproteins.
• three classes of glycoproteins

❶ Glycoproteins
• In class, the protein constituent is the largest
component by weight.
• a variety of biochemical roles: components of
cell membranes (cell adhesion and the binding
of sperm to eggs).
❷ Proteoglycans
• The protein component is conjugated to a particular
type of polysaccharide called a glycosaminoglycan
• Carbohydrates are the largest component
• Proteoglycans function as structural components and
lubricants.

❸ Mucins or mucoproteins
• Are predominantly carbohydrate.
• N -Acetylgalactosamine is usually the
carbohydrate bound to the protein in mucins.
• Mucins, a key component of mucus, serve as
lubricants.

Each blood group is designated by the presence of one of the

! three different carbohydrates, termed A, B, or O, attached to
glycoproteins and glycolipids on the surfaces of red blood cells 3
❶ Glycoproteins
Sugars in glycoproteins are attached either to the amide nitrogen atom in the side chain of asparagine (termed an N- linkage)
or to the oxygen atom in the side chain of serine or threonine (termed an O- linkage)

serine or
asparagine threonine

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Erythropoietin (EPO) = glycoprotein hormone, present in the blood serum
that improved treatment for anemia, particularly that induced by cancer
chemotherapy

• secreted by the kidneys, stimulates the production of red blood cells

• 165 amino acids
• N -glycosylated at 3 Asn residues and O -glycosylated on a Ser residue
• mature EPO is 40% carbohydrate by weight,
• glycosylation inproves stability
• Unglycosylated protein has only about 10% of the bioactivity of the
glycosylated
• endurance athletes have used recombinant human EPO to increase the
red-blood-cell count and hence their oxygen-carrying capacity.
• Drug-testing laboratories are able to distinguish some forms of prohibited
EPO
human recombinant EPO from natural EPO in athletes by detecting
differences in their glycosylation patterns

Lance Armstrong
5
❷ Proteoglycans = proteins attached to glycosaminoglycans (aprox 95% of the weight) so it resembles a polysaccharide
more than a protein
• lubricants, structural components in connective tissue
• The properties of proteoglycans are determined by the glycosaminoglycan component.
• many glycosaminoglycans contain a derivative of an amino sugar - glucosamine or galactosamine

Chondroitin 6-sulfate Keratan sulfate Heparin

Anticoagulant

Dermatan sulfate Hyaluronate

Major glycosaminoglycans 6
Proteoglycans in cartilage
structure
! 2 importants components of cartilage: proteoglycan aggrecan + protein collagen
Shock absorber Strength
• 2397 amino acids
• The protein has three
globular domainswhere
keratan sulfate and
chondroitin sulfate are
attached
• water is bound to the negative charges of glycosaminoglycans
and serves a cushion
• when pressure is exerted, as when the foot hits the ground
while walking, water is squeezed from the glycosaminoglycan,
cushioning the impact.
• When the pressure is released, the water rebinds.
• Osteoarthritis - water is lost from proteoglycan with aging
or the proteolytic degradation of aggrecan and collagen in
the cartilage. 7
❸ Mucins or mucoproteins
The protein component is extensively glycosylated at serine or
threonine residues by N -acetylgalactosamine

N -acetylgalactosamine

Mucins form large polymeric structures and are common in mucous

secretions (E.g. saliva)

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 3.2.2.2. Lipoproteins VLDL (very low density
lipoproteins) have very high
• prosthetic component = fatty acids, density small
triglycerides, lecithins, cephalins, • transport the triglycerides
cholesterol, fat-soluble vitamins. LDL (low density lipoprotein) synthesized in the liver to
• are part of membrane structure, or represents the cholesterol the extra hepatic tissues Chilomicrons have low
are found in plasma and have the role transportor for tissues density and large size
of transporting plasma lipids. • a risk factor in atherosclerosis • transport food lipids
• Although they have a high lipid
content (sometimes 90%), they are
still water soluble, which is why it is
admitted that the protein part is
located on the outside, and the lipid
part on the inside
• Lipoproteins are classified by density
(the higher the lipid content, the
lower the density)

HDL (high density lipoproteins) have high

density
• transport of cholesterol from tissues to
the liver where it is transformed 9
Chromoproteins

10
 1. Hemoglobin vs Myoglobin - General considerations

Myoglobin is located in muscle

Hemoglobin is contained in red
• facilitates the diffusion of
blood cells
oxygen through the cell for the
• Carries oxygen from the lungs
generation of cellular energy
to the tissues
• provides a reserve supply of
• transport of carbon dioxide
oxygen available in time of
and hydrogen ions back to the
need.
lungs.

Hb is a remarkably efficient oxygen carrier, able to use as much

as 90% of its potential oxygen-carrying capacity while under
similar conditions, Mb is able to use only 7% of its potential
capacity
WHY?
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 Hemoglobin vs Myoglobin

Myoglobin has a single polypeptide chain, whereas hemoglobin comprises four polypeptide chains.
The four chains in hemoglobin bind oxygen cooperatively, meaning that the binding of oxygen to a
site in one chain increases the likelihood that the remaining chains will bind oxygen.

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2. Heme structure
HEME
• the organic part - protoporphyrin IX (1,3,5,8
tetramethyl-2,4 divinyl-6,7 dipropionyl porphyrin)
consists of 4 pyrrole nuclei linked by methyn bridges.
• Substituents: methyl (4), vinyl (2) and propionyl (2);

• an Fe2+ (ferrous) ion in the center of protoporphyrin IX

Porphyrin bound to the nitrogen atoms of the pyrrole nucleus by
two covalent bonds and two coordinative bonds.
• The iron ion forms two other bonds (the 5th and 6th
iron coordination) with 2 histidine residues (proximal
and distal histidine) from the globin structure.

methyl
methylene The heme group gives
methine Fe-Protoporphyrin IX ! muscle and blood their
distinctive red color
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3. Oxygen binding to Myoglobin and Hemoglobin
Oxymyoglobin/oxyhemoglobin

N (distal
O2 histidine)

(proximal N
histidine)

N (distal histidine)

(proximal N His
histidine)

Deoxymyoglobin/deoxyhemoglobin 14
 3.1. Factors that influence oxygen binding to hemoglobin
3.1.1 pO2

• Mb curve rises sharply as pO2 increases
and then levels off.
• Half-saturation (p50) is at the relatively low
value of 2 torr (mm Hg), indicating that
oxygen binds with high affinity to
myoglobin.
Oxygen saturation
• Hb curve does not look like a simple binding curve such as that
for myoglobin; instead, it resembles an “S” (sigmoid)
• P50 = 26 torr indicated that oxygen binding is significantly
weaker than that for myoglobin
• Sigmoid shape suggests that the binding of oxygen at one site
within the hemoglobin tetramer increases the likelihood that
oxygen binds at the remaining unoccupied sites and vice versa
the unloading of oxygen at one heme facilitates the unloading
of oxygen at the others = cooperative binding

P50 = 2 torr

p 50 = represents the oxygen presure when 50% of

the molecule is saturated with oxygen
p 50 = 26 torr for Hb
p 50 = 2 torr for Mb 15
 3.1.2. Cooperative binding of oxygen to Hb - physiological significance

1. Oxygen must be transported in the blood from the

lungs to the tissues

2. In the lungs pO2 is relatively high (pO2=100 torr)

while in the tissues partial pressure of oxygen is much
lower (pO2=20 torr).

3. In the lungs, hemoglobin becomes nearly saturated

with oxygen ( 98% of the oxygen-binding sites are
occupied)

4. In the tissues Hb releases O2 (oxygen saturation

drops to 32%), so 98 – 32 = 66% of the oxygen is
*Normal atmospheric pressure is defined as 1
unloaded in the tissues atmosphere. 1 atm = 14.6956 psi = 760 torr.

Mb is 98% saturated with oxygen in the lungs and it remains
91% saturated in the tissues, so only 98 - 91 = 7% of the sites
Mb
would contribute to oxygen transport; myoglobin binds oxygen
too tightly to be useful in oxygen transport.
The cooperative binding of oxygen by Hb
requires that the binding of oxygen at one site
in the hemoglobin tetramer influence the
oxygen binding properties at the other sites.
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3.1.3. 2,3-Bisphosphoglycerate (2,3 BPG)

• 2,3 BPG influences oxygen binding to HB

• 2,3 BPG is a highly anionic compound present in red blood cells at
approximately the same concentration as that of Hb
• without 2,3-BPG, hemoglobin would release only 8% of oxygen in the tissues
• 2,3 BPG binds to Hb in a cavity in the center of the tetramer and decreases its
affinity for O2, resulting in O2 release from oxyHb.

Physiological consequences of BPG binding to Hb

• Protein part (globin) in human fetuses differs from adults

• fetal hemoglobin tetramers include two alpha chains and two gamma chains
• the gamma chain resembles adult beta chain (72% identical) but one Ser
residue is substituted with His, so 2,3 BPG affinity for fetal Hb is reduced
• Consequently, the oxygen binding affinity of fetal Hb is higher than that of
maternal (adult) HB => oxygen to be effectively transferred from maternal to
fetal red blood cells. 17
3.1.4. Hydrogen Ions Promote the Release of Oxygen: The Bohr Effect

• Rapidly metabolizing tissues, such as contracting muscle, generate large amounts of hydrogen ions and
carbon dioxide

1 2
In the tissues, where CO2 and lactic acid are In the lungs, where CO2 is
formed from metabolism, it results in a released, there is a slight increase
decrease in pH (pH=7.2), which favors O2 in pH (pH=7.4), Hb binds more O2,
release, until Hb reaches a saturation of tends to saturate with O2.
maximum 60%.

The dependence of the Hb affinity for O2 as a function of pH represents the Bohr effect

H+ Hb + O2 ↔ Hb O2 + H+

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4. Carbon monoxide binding to Hemoglobin

• Combination of Hb with CO generates carboxyhemoglobin

• The affinity of Hb for CO is 200 times higher than for O2.
• If 50% of the erythrocyte Hb is in the form of HbCO, dizziness, headache occur, and if the process
continues to increase, even deadly poisoning can occur.

Hb + CO Hb CO

• reaction is reversible, but the balance is shifted to the right, even at low CO concentrations large amounts
of HbCO are formed
• In CO intoxications oxygen therapy is applied at high pressure.
• In smokers the HbCO concentration can reach up to 10%.

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5. Carbon dioxide transport

The function of CO2 transport from tissues to the

lungs is accomplished by binding CO2, but not
through the heme, but by a -NH2 group in the
globin structure, with carbaminohemoglobin
formation:
Hb-NH2 + CO2 →Hb-NH-COOH + H+
carbaminohemoglobin
≈14 %

In red blood cells carbon dioxide is reversibly

transformed under the action of carbonic
anhydrase: In the lungs, this process is
reversed: HCO3- is converted
carbonic anhydrase
CO2 + H2O H+ + HCO3- back into CO2 and exhaled
H2CO3

carbonic acid dissociates to form bicarbonate

ion (HCO3-) and and H+, resulting in a drop in pH
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(Bohr Effect) = oxygen release in the tissues
6. Pathological aspects
Thalassemia is caused by the loss or
• A single amino acid substitution
Sickle-cell anemia substantial reduction of a single hemoglobin
(replacement of a glutamate residue
chain
with valine in position 6) in the beta
• Results in low levels of functional
chain of Hb produces hemoglobin S
hemoglobin and a decreased production
(HbS).
of red blood cells, which may lead to
• HbS forms large fibrous aggregates
anemia, fatigue, pale skin, and spleen
that extend across the red blood
and liver malfunction.
cells, distorting them so that they
• In alpha -thalassemia, the alpha chain of
clog small capillaries and impair
hemoglobin is not produced in sufficient
blood flow.
quantity so Hb tetramers contain only the
• red cells from sickle cell patients are
beta chain (HbH), bind oxygen with high
more adherent to the walls of blood
Sickled red blood cells affinity and no cooperativity.
vessels increasing the risk of capillary
• In beta -thalassemia, the beta chain of Hb
occlusion (painful swelling of the
is not produced in sufficient quantity. In
extremities and a higher risk of
the absence of beta chains, the alpha
stroke or bacterial infection
chains form insoluble aggregates that
• The sickled red cells also do not
precipitate inside immature red blood
remain in circulation as long as
cells. The loss of red blood cells results in
normal cells do, leading to anemia.
anemia. 21
Nucleoproteins

22
Nucleoproteins = hetero-proteins that are found in the cell nucleus or mitochondria consisting of
a protein part and a prosthetic group (nucleic acid)
Replication
Transcription Translation
DNA RNA PROTEIN

1. General considerations

Nucleic acids are produced by polycondensation of the nucleotides, so they are polynucleotides.
The binding of the protein part to the prosthetic part is made, in particular, by ionic but also covalent
bonds.

RNA DNA

pentose nitrogenous bases phosphate residue pentose nitrogenous bases phosphate residue
ribose Adenine deoxyribose Adenine
Guanine Guanine
Cytosine Cytosine
Uracil Thymine
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2. Pentose 3. Nitrogenous bases

Purine Adenine, A Guanine, G

Ribose Deoxyribose

Pyrimidine

Cytosine, C Uracil, U Thymine, T

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4. Nucleosides = N - glycosides of purine or pyrimidine bases with ribose, respectively deoxyribose.

An N-glycosidic bond is formed between hydrogen at position 9 of the purine bases or position 1 of the pyrimidine bases
and –OH group in position 1’ of pentose

Adenosine Guanosine Citidine Uridine Deoxythymidine

25
5. Nucleotides = are phosphoric esters of nucleosides, so they consist of a nitrogenous base + pentose + phosphate residue

Adenylic acid, Adenosine monophosphate, AMP

Guanylic acid, Guanosine monophosphate, GMP
Cytydylic acid, Cytidine monophosphate, CMP
Uridylic acid, Uridine monophosphate, UMP
Deoxythymidylic acid, Deoxythymidine monophosphate, dTMP

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6. Nucleic acids = are, with few exceptions, linear polymers of nucleotides whose phosphate groups bridge the 3’ and 5’
positions of successive sugar residues
• The phosphate groups, at physiological pH’s, are in the form
of anions.
• Polynucleotides have directionality, that is, each has a 3’ end
5’ end (the end whose C3’ atom is not linked to a neighboring
nucleotide) and a 5’ end (the end whose C5’ atom is not
linked to a neighboring nucleotide).

6.1. DNA
• DNA has a double-helical structure
• DNA has equal numbers of adenine and thymine residues (A = T) and equal
numbers of guanine and cytosine residues (G = C).

3’ end
27
Chemical structure of a nucleic acid
 Watson and Crick developed the double helical DNA
conformation model, for which they received the Nobel Prize,
in 1962. Following this model, the DNA has a double helix
structure, consisting of two antiparallel polynucleotide chains.
The outer diameter of the double helix is 20 Å (2 nm), the spin
pitch is 34 Å (3.4 nm), with the bases facing inwards.

• each polynucleotide chain has two ends: one with the 5‘ end –OH group is free
and one with the 3'end –OH group free.
• By convention, a polynucleotide chain is written with the 5 'end on the left and
the 3' on the right.
• E.g. the sequence of AGCAC means that adenine has the 5 '-OH group not
bound to another nucleotide, while the cytosine has the free 3' -OH group.
• Two polynucleotide chains are antiparallel, when one runs in the 5 '→ 3'
direction and the other in the 3 ' → 5' direction.

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• To fit in a small place (nucleus) always next to a purine base on one
chain, a pyrimidine base will be placed on the other chain and vice
versa.
• This three-dimensional conformation is stabilized by hydrogen bonds,
which are formed between the complementary bases that are facing
each other:
A === T is bound by up to 2 hydrogen bonds;
G≡≡≡C is bound by a maximum of 3 hydrogen bonds.
• Principle of complementarity of the nitrogen bases = the two
polynucleotide chains of the double helix are not identical, but
complementary, in the sense that the adenine on one chain will
always correspond to the thymine on the other chain, and to the
guanine - cytosine.
• Genetic information regarding protein biosynthesis is given by the
sequence of the nitrogen bases in the DNA strands.
• Due to structural complementarity, the ratio A + G / T + C =1 and the
ratio A + T / G + C varies from one DNA species to another.

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6.2. RNA

• RNA molecules are single stranded, but, on certain portions, where the complementarity of bases A == U, G≡≡C allows,
double helical structures are organized
• The content of adenine ≠ uracil and guanine ≠ cytosine excludes the existence of a complementary double-stranded
structure.
• The portions with non-complementary bases are expelled as loops outside the double helix areas, giving the molecule a
"hairpin" appearance.
tRNA
• consists of aprox. 76 nucleotides and
mRNAs are essentially a series of consecutive 3- contains a trinucleotide sequence, its
nucleotide segments known as codons, each of anticodon, which is complementary to the
which specifies a particular amino acid. codon(s)
• An amino acid is covalently linked to the 3’
end of its corresponding tRNA to form an
aminoacyl-tRNA
• During translation, the mRNA is passed
through the ribosome such that each codon,
in turn, binds its corresponding
aminoacyltRNA

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Nucleoside triphosphates.
Biochemical energy concepts

31
 ATP = energy currency of our body • A part of the energy produced by oxidation of food is transformed into this
highly accessible molecule
• ATP also acts as the free-energy donor in most energy-requiring processes
such as motion, active transport and biosynthesis.
Macroenergetic bonds

• In most metabolic processes, the energy released from

Macroergic nucleotides clasification
• purine nucleotides: adenosine di- and triphosphate, ADP
and ATP, guanosine di- and triphosphate, GDP and GTP;
• pyrimidine nucleotides: cytidine di- and triphosphate,
CDP and CTP, uridin di- and triphosphate, UDP and UTP,
trimidine di- and triphosphate, TDP and TTP.
the cleavage of a macroergic bond in ATP is sufficient, in
this case the ATP cleaves to ADP and a phosphate
residue H3PO4 (Pi) and a quantity of energy equivalent
to 7.3 kcal / mol is released.
• There are also reactions in which more than 7.3 kcal is
needed, in which case both ATP macro-hydrolyzes are
hydrolyzed with AMP formation and a pyrophosphate
residue H4P2O7 (PPi).
• Pyrophosphate is then cleaved under the action of
pyrophosphatase into two Pi phosphate residues,
leading to the release of an additional 8 kcal energy.
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 Legături
Macroenergetic
macroergice
bonds ATP

H2O

H2O
H3PO4 +
7,3 kcal/mol

H4P2O7 + 7,3 kcal/mol

Pirophosphatase
H2O Pirofosfataza

2 H3PO4 + 8 kcal/mol ADP

AMP
33
 Reactions that require ATP

❶ enzymatic phosphorylation of glucose at Glucose-6-Phosphate requires less than 7.3 kcal / mol, which is why hydrolysis
of ATP → ADP + PI is sufficient;

Glucokinase
1

ATP ADP
Glucoza G-6-P

❷ more than is required for the activation of fatty acids in acyl-CoA 7.3 kcal / mol, for this reason ATP is cleaved into AMP
and PPi, pyrophosphate is rapidly cleaved into 2 Pi with a higher energy release.

❸ UTP is used in glucose metabolism to activate glucose; CTP is involved in lipid biosynthesis; GTP participates in protein
biosynthesis together with ATP.

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