Biochemistry of Lens

Chemical Composition of the Lens

Membranes
lens-fiber plasma membranes are very stable and very rigid.
High content of saturated fatty acids, high cholesterol to phospholipid ratio, high
concentration of sphingomyelin tight packing and low fluidity of the
membrane.
Lens Proteins
33% of its wet weight probably the highest protein content of any tissue.
Lens proteins are commonly divided into 2 groups based on water solubility
Lens Proteins
Crystallin Proteins
Providing the transparency and refractile properties.
Have 2 attributes:
Very stable structures
Remain soluble under conditions of high protein concentration without
forming large aggregates, which would be lightscattering
Crystallins can be divided into 2 groups:
-crystallin and the ,-crystallin family present in all vertebrate lenses.
Taxon-specific crystallins is present only in phylogenetically restricted
groups of species.
-Crystallin
The largest of the crystallins, represent about one-third of the lens
proteins by mass.
Has a native molecular mass ranging between 600 and 800 kD
Composed of 2 subunits: A and B mass of approximately 20 kD,
nearly 55% identical in sequence.
Inhibit denaturation and insolubilization of the other crystallins :
-Crystallin complexes bind to partially denatured proteins prevent
further denaturation and aggregation
Antiaggregative function synthesis of new protein is impossible
maintenance of transparency in the fibers of the lens nucleus
Zinc ions enhance the chaperone function and stability of -crystallin.
,-Crystallins
Related members of the same protein superfamily.
Might have arisen from double duplication and fusion of a gene for a 40-residue
polypeptide.
Subdivided into 2 groups :
-crystallins
Complex group of oligomers
composed of polypeptides
Encoded by 7 genes.
Molecular masses ranging from 23 to
32 kDa.
The individual polypeptides associate
with each other, forming dimers and
higher-order complexes in their native
state.
can be separated into H ( high-
molecular-mass) and L ( low-
molecular-mass) fractions
-crystallins
Monomeric proteins
In humans: encoded by 4 genes.
Molecular mass 20 kDa or less (smallest
crystallins).
The native -crystallins do not associate with
each other or with other proteins lowest
molecular mass of the crystalline fractions.
Most expression occurs early in development
most concentrated in the nuclear region of
the lens.
Compact and symmetric structures tend to
be highly concentrated in aged, hard lenses,
with little to no accommodative ability.
Taxon-specific crystallins
Most taxon-specific crystallins are oxidoreductases bind pyridine
nucleotides
Presence in the lens significantly increases the concentration of the
bound nucleotides.
Reduced nucleotides absorb ultraviolet (UV) light and may protect the
retina from UV-induced oxidation.
PENINGKATAN PROTEIN TIDAK LARUT
AIR
HIPOTESA :
Protein Agregasi partikel
tidak larut dalam air sgt besar

Kekeruhan lensa Memecah cahaya

Membrane Structural Proteins and Cytoskeletal Proteins

Cytoskeletal and membrane proteins

Several structural proteins can be solubilized only in the presence of
chaotropic agents or detergents:
cytoskeletal elements actin (actin filaments),
vimentin (intermediated filaments),
tubulin (microtubules),
2 additional proteins: filensin and phakinin found only in lens-fiber cells
and compose a cytoskeletal structure
Water-insoluble fraction of lens proteins can be further divided into 2
fractions: 8 M Urea Soluble AND 8m Urea Insoluble
8 M Urea Soluble
Most cytoskeletal protein provide the structural framework of the lens
cells.
Microfilaments and microtubules are similar to those in other cell types.
2 types of intermediate filaments that are unusual:
one class made from the protein vimentin (not usually found in epithelial cells);
the beaded filaments : composed of the proteins phakinin and filensin, specific to
the lens.
Genetic disruption of beaded filaments structure disruption of the
structure of the lens fiber cells formation of cataract.
8 M Urea Insoluble fraction
Known as the major intrinsic protein (MIP; also known as aquaporin 0)
MIP is expressed only in lens-fiber cells
Functions as a water channel.
Has a molecular mass of 28 kDa
Associated with fiber cell plasma membranes, makes up nearly 50% of the membrane
proteins
MIP first appears in the lens just as the fibers begin to elongate with age undergoes
proteolytic cleavage forming a protein fragment with a molecular mass of 22 kDa.
At 2030 years of age the relative proportions become about equal.
Over time, the protein with molecular mass of 22 kDa predominates in the lens nucleus.
Increase of Water-Insoluble Proteins With Age

As the lens ages proteins aggregate form very large particles become
water-insoluble and scatter light, increasing the opacity of the lens.
The water-insoluble protein fraction increases with age, even if the lens remains
relatively transparent.
Conversion of the water-soluble proteins into water insoluble proteins appears to
be a natural process in lens fiber maturation, but it may occur more quickly in
cataractous lenses.
Brunescent cataracts the increase in the amount of water-insoluble protein
correlates well with the degree of opacification.
Increase of Water-Insoluble Proteins With Age

Associated oxidative changes occur : protein-to-protein and protein-to-

glutathione disulfide bond formation decreased levels of the reduced
form of glutathione and increased levels of glutathione disulfide (oxidized
glutathione) in the cytoplasm of the nuclear fiber cells.
Depletion of the reduced form of glutathione accelerates protein
crosslinking, protein aggregation, and light scattering.
In addition to the increased formation of disulfide bonds, nuclear proteins
are highly crosslinked by nondisulfide bonds contains yellow-to-brown
pigments that are found in higher concentration in nuclear cataracts.
Increased fluorescence is generated by the nondisulfide crosslinks that
form in brunescent nuclear cataracts.
Posttranslational modifications to lens proteins
Proteins of the lens are probably the longest-lived
The oldest ones (center of the lens nucleus) are synthesized before birth.
Structurally modified in various ways:
oxidation of sulfur and aromatic residue side chains,
inter- and intrapolypeptide crosslinking,
glycation,
racemization,
phosphorylation,
deamidation,
carbamylation.
 As the proteins age oxidative modifications accumulate
crosslinking of crystallin polypeptides, alterations in fluorescent
properties, and an increase in protein-associated pigmentation.
Formation of disulfide crosslinks in the proteins of the lens nuclear
region is associated with the formation of protein aggregates, light
scattering, and cataract
Carbohydrate Metabolism
Lens
Goals Transparency

Glucose Simple Diffusion

Facilitated Diffusion Lens
Most of the glucose transported into the lens is phosphorylated
to glucose-6-phosphate (G6P) by the enzyme hexokinase.
This reaction is 70100 times slower than that of other enzymes
involved in lens glycolysis and is, therefore, rate-limited in the
lens.
Carbohydrate Metabolism
2 METABOLIC PATHWAY ANAEROBIC GLYCOLYSIS

G6P
HEXOSE MONOPHOSPHATE
(HMP SHUNT)

Jalur yang lebih aktif adalah glikolisis anaerobik, yang menyediakan ikatan fosfat energi tinggi
terbanyak yang dibutuhkan untuk metabolisme lensa.
Langkah dengan kecepatan yang terbatas pada jalur glikolisis sendiri ada pada tahap enzim
fosfofruktokinase yang diatur melalui umpan balik oleh produk metabolik dari jalur glikolisis.
Jalur ini lebih sedikit efisiensinya dibandingkan dengan glikolisis aerobik yang menghasilkan 36
molekul ATP dari setiap molekul glukosa yang dimetabolisme dalam siklus asam sitrat
(metabolisme oksidatif).
Karena tekanan oksigen yang rendah dalam lensa, hanya sekitar 3% dari glukosa lensa yang
melewati siklus asam sitrat Krebs untuk memproduksi ATP; bagaimana pun, walau hanya dengan
metabolisme aerobik yang rendah, dapat menghasilkan 25% dari ATP lensa.
 Lensa tidak tergantung pada O2, lensa dapat menjaga metabolisme
normal dalam lingkungan nitrogen.
Dengan diberikan sejumlah glukosa, lensa in vitro yang anoksik tetap
jernih dan utuh, memiliki kadar normal dari ATP serta
mempertahankan aktivitas pompa asam amino dan ion. Namun,
ketika glukosa menurun, lensa tidak dapat mempertahankan fungsi-
fungsi ini dan menjadi keruh pada beberapa jam sekalipun terdapat
oksigen.
 Jalur yang kurang aktif untuk utilisasi G6P dalam
lensa adalah heksosa monofosfat shunt (HMP
Shunt), yang dikenal juga dengan istilah jalur
pentosa monofosfat.
5% glukosa lensa dimetabolisme melalui jalur ini
sekalipun jalur ini distimulasi peningkatan kadar
glukosa.
Aktifitas HMP Shunt lebih tinggi pada lensa
dibandingkan dengan jaringan lain. Seperti pada
jaringan lain, dapat menghasilkan NADPH (sebuah
bentuk tereduksi dari nicotinamide-adenine
dinucleotide phosphate (NADP)) untuk biosintesis
asam lemak dan biosintesis ribosa untuk
nukleotida.
Dihasilkan pula NADPH untuk aktifitas glutation
reduktase dan aldose reduktase dalam lensa.
Produk karbohidrat dari HMP shunt memasuki
jalur glikolisis dan dimetabolisme menjadi laktat.
Aldose reduktase adalah enzim kunci pada jalur
lain metabolisme karbohidrat pada lensa yaitu
jalur sorbitol.
Mekanisme Umpan Balik
Ketika kadar glukosa meningkat dalam lensa sebagaimana terjadi pada
keadaan hiperglikemia, jalur sorbitol teraktifasi lebih daripada glikolisis dan
terjadi akumulasi dari sorbitol.
Sorbitol dimetabolisme menjadi fruktosa oleh enzim polyol dehidrogenase.
Sayangnya enzim ini memiliki affinitas yang rendah yang berarti sorbitol
akan terakumulasi sebelum mengalami metabolisme labih lanjut.
Karakteristik ini, dikombinasikan dengan kurangnya permeabilitas lensa
terhadap sorbitol berakhir dengan retensi sorbitol dalam lensa.
Sejalan dengan sorbitol, fruktosa juga terbentuk pada lensa dengan kadar
tinggi glukosa. Bersamaan, kedua gula tersebut meningkatkan tekanan
osmotik di dalam lensa dan menarik air. Pada mulanya pompa tergantung
energi pada lensa mampu mengkompensasi, tetapi akhirnya kemampuan
tersebut terlewati. Hasilnya adalah pembengkakan serat, rusaknya
arsitektur sitoskeletal normal dan kekeruhan lensa.
KERUSAKAN OKSIDATIF DAN MEKANISME
PROTEKSI
Aktivitas metabolik
1. Radikal bebas
2. Agen eksterna (energi radiasi)
Radikal Bebas relatif tinggi kerusakan serat lensa
Peroksidasi (dlm serat plasma/mb.plasma)
kekeruhan lensa
DNA reversible
irreversible
Protein atau mb lipid (korteks lensa)
MEKANISME PROTEKSI
Poteksi Radikal Bebas

Enzim Vitamin C dan E

Pemecah radikal bebas

kerusakan oksidatif

1. Glutathione peroksidase
2. Catalase
3. Superoxide dismutase
Sumber
AAO The Eye M.D. Association. 2016-2017. Fundamentals and Principales
of Ophthalmology, Lens and Cataract, BCSC Section 2 & 11
Terima kasih, mohon saran dan bimbingan