Biochemistry for Dental Students

1. Dental Biochemistry 2 – (Lec. 4)
Connective tissue and
muscle protein

2. Collagen
• The major structural protein found in connective tissue is
the collagen.
• Collagen is a Greek word which means the substance to
produce glue.
• It is the most abundant protein in the body.
• About 25-30% of the total weight of protein in the body is
collagen.
• It serves to hold together the cells in the tissues.
• It is the major fibrous element of tissues like bone, teeth,
tendons, cartilage and blood vessels.
• When a solution of collagen is boiled, the viscosity of the
solution decreases, which indicates that the native rod like
structure is altered and a protein, with random coil structure
results. It is then called gelatin.

5. Structure of Collagen
• The tropocollagen is made up of three polypeptide
chains.
• There are 6 types of collagen, out of which type I is
the most abundant form; it contains 2 chains of
alpha-1 and one chain of alpha-2.
• Each polypeptide chain of collagen has about 1000
amino acid residues.
• The amino acid composition of collagen is quite
unique.

6. • About 33% of the amino acids is glycine, that is,
every third residue is glycine.
• The repetitive amino acid sequence may be
represented as Gly – X – Y – Gly – X - Y; where X
and Y are other amino acids, most commonly
proline, hydroxyproline and hydroxy lysine are
found in fairly large proportions in collagen.
• The hydroxylated amino acid residues are of
special functional significance.

8. Synthesis of Collagen
• The collagen is synthesized by fibroblasts
intracellularly, as a large precursor,
called procollagen.
• It is then secreted.
• The extracellular procollagen is cleaved
by specific peptidases to form
tropocollagen.

10. Hydroxylation of Proline and Lysine
• The hydroxylation of proline and lysine
residues of collagen is a post-translational
modification taking place intracellularly.
• Prolyl hydroxylase and lysyl hydroxylase
enzymes contain ferrous iron at the active
site and require a reducing agent like
ascorbic acid.
• So, vitamin C deficiency leads to poor
hydroxylation.
• It is the major biochemical defect in scurvy.

11. Triple Standed Helix
• The collagen is a rod like structure.
• Each of the 3 polypeptide chains is held in a helical
conformation by winding around each other.
• The resulting cable is made in a manner that 3.3
amino acid residues make one turn and each turn is
separated by 2.9 A°.
• The three strands are hydrogen bonded to each other.
• Glycine, because of its small size can fit into.
• For the same reason, glycine also produces a shallow
groove into which other polypeptide strands are
interwined.

12. Quarter Staggered Arrangement
• The Tropocollagen molecules are arranged
in a ‘quarter staggered array’ to form
collagen fibers
(Molecules in each row separated by 400 A°
and adjacent rows by 680A°).
• The structure repeats after fifth row.
• Thus the collagen fiber has triple stranded,
quarter staggered arrangement.
• This arrangement helps in mineralization.

14. Cross Linked in Collagen Fibers
• The collagen fibers are strengthened by covalent
cross-links between lysine and hydroxy lysine
residues.
• The cross links are formed by lysyl oxidase.
• It is a copper containing enzyme, the copper ion being
located at its active site.
• In copper deficiency, collagen synthesis is abnormal.
• The older the collagen, the more the extent of cross
linkages.
• The process continues, especially in old age, so that the
skin, blood vessels and other tissues become less
elastic and more stiff, contributing a great extent to the
medical problems of the old people.

15. Function of Collagen
1. To give support to organs.
2. To provide alignment of cells, so that
cell anchoring is possible. This in
turn, helps in proliferation and
differentiation of cells.
3. In blood vessels, if collagen is
exposed, platelets adhere and
thrombus formation is initiated.

16. Abnormalities in Collagen
1. Osteogenesis imperfecta:
• It is inherited as a dominant trait.
• It is the result of a mutation which results in the
replacement of a single glycine residue by cysteine.
• This change disrupts the triple helix near the
carboxy terminus, hence the polypeptide becomes
excessively glycosylated and hydroxylated.
• So, unfolding of the helix takes place and fibrillar
array cannot be formed.
• This results in brittle bones leading to multiple
fractures and skeletal deformities.

17. 2. Ehlers-Danlos syndrome:
• It is due to defective collagen formation.
• It is characterized by loose skin, hypermobile and
lax joints.
3. Deficiency of Ascorbic Acid:
• It is characterized by defective hydroxylation of
collagen.
• The collagen formed is weak, leading to fragility
of blood vessels, poor wound healing, bleeding
gum, etc.

18. 4. Copper deficiency:
• Copper deficiency blocks the lysyl
oxidase, resulting in reduced
formation of cross linking.
• The elastic nature of elastin fibers are
due to these different cross links.

19. Elastin
• Elastin is a protein found in connective
tissue and is the major component of
elastic fibers.
• The elastic fibers can stretch and then
resume their original length.
• They have high tensile strength.
• They are found in the ligaments as well
as in the walls of the blood vessels,
especially large vessels like aorta.

20. Keratins
• Keratine are fibrous proteins present in hair,
skin an nail, horn, hoof, etc.
• They mainly have the alpha helical structure.
• Each fibril has 3 polypeptide chains and each
bundle has about 10-12 fibrils.
• The matrix has cysteine-rich polypeptide chains
which are held together by disulfide bonds.
• The more the number of disulfide bonds, the
harder the keratin is.

21. Muscle Proteins
• Striated muscle is made up of multinucleated
cells bound by plasma membrane called
Sarcolemma.
• Each muscle cell contains myofibrils about 1mm
in diameter.
• The functional unit of a myofibril is a sarcomere.
• The dark A bands and light I bands alternate
regularly.

22. • These bands are formed by variable combination of
thick and thin filaments.
• The thick filament is primarily myosin and thin
filament contains actin, tropomyosin and
troponin.
• Thick and thin filaments slide past each other during
the muscle contraction, so that the muscle shortens
by as much as a third of its original length.
• However the length of the thick and thin filaments
do not change during muscle contraction.

26. Myosin
• Myosin molecules are large (about
540 KD), each with 6 polypeptide
chains.

27. Actin
• It is the major protein of the thin filaments.
• It is a monomeric protein often referred to as G-
actin due to its globular shape.
• It can polymerize into a fibrous form, called F-actin,
which is a helix of actin monomer.
• The muscle contraction results from interaction of
actin and myosin, to form actomyosin, with energy
provided by ATP.
• When the two thin filaments that bind the cross
bridges of a thick filament are drawn towards each
other, the distance between Z lines becomes
shorter.

29. • This could result in the process of contraction of
muscle fibers.
• This needs energy from hydrolysis of ATP, effected by
the ATPase activity of myosin.
• The contractile force is generated by conformational
changes, leading to cyclic formation and dissociation
of actin and S1 heads of myosin.
• There is a reversible attachment and detachment of
myosin S1 head to actin.
• This is due to the hinge like movements between the
domains of myosin.
• The action of calcium is brought about by 2 proteins,
troponin complex and tropomyosin located in the thin
filament.

30. • The troponin complex has 3 different
polypeptide chains.
• Out of this, troponin-C (TnC) binds
calcium.
• Troponin-I (TnI), binds to actin and
inhibits binding of actin to myosin.
• Troponin I is a marker for myocardial
infarction.
• Its level in serum is increased within 4
hours of myocardial infarction.

31. Troponin-T (TnT) binds to tropomyosin.
• Two isomers of cardiac TnT, called TnT1 and TnT2
are present in adult human cardiac tissue.
• Serum levels of TnT2 increases within 4 hours of
myocardial infarction, and remains high for up to
14 days.
• The TnT2 is 100% sensitive index for myocardial
infarction.
• The reservoir of high energy phosphateis skeletal
muscle is creatine phosphate.
• The reaction (Lohman’s reaction).
CK
• Creatine phosphate + ADP -------→ ATP + Creatine

32. • During muscle contraction, the ATP level remains
high as long as creatine phosphate is present.
• But following contractile activity, the level of ADP
and Pi rises.
• The reduced energy charge of active muscle
stimulates glycogen breakdown, glycolysis, TCA
cycle and oxidative phosphorylation, so that energy
is derived from aerobic metabolism.
• Hence, only aerobic exercise is useful for weight
control.