CHAPTER 3 ophthalmologyebooks.com
Biochemistry and Physiology
See BCSC Section 2, Fundamentals and Principles of Ophthalmology, for
additional discussion of several of the topics discussed in this chapter.
Molecular Biology
Crystallin Proteins
‘The human lens has a protein concentration of 33% of its wet weight. which is at
least twice that of most tissues. Lens proteins are commonly divided into 2 groups,
based on water solubility (Fig 3-1). The water-soluble fraction of the young lens
accounts for approximately 80% of lens proteins: aging-related changes result in an
increase in the percentage of water-insoluble protein. The water-soluble portion
consists mainly of a group of proteins called crystallins. The crystallins can be
divided into 2 major groups. o-crystallins and B.y-crystallins.
Lens Proteins
Water soluble
{Intracellular proteins) Water insoluble
orCrystallins Crystalis Urea soluble Urea insoluble
(most cytoskeletal (most lens fiber cell membrane
proteins) proteins; includes major
intrinsic protein [MIP])
Figure 3-1 Overview of lens proteins.The nrea-insoluble fraction of the young lens contains the plasma membranes
of the lens fiber cells. Several proteins are associated with these fiber cell plasma
membranes. One makes up nearly 50% of the membrane proteins and is known as
the major intrinsic protein (MIP; also known as aquaporin 0), a member of a
class of proteins called aquaporins. Other members of the aquaporin family are
found throughout the body, where they serve predominantly as water channels.
MIP first appears in the Jens just as the fibers begin to elongate. With age, this
protein, which has a molecular mass of 28 kDa, undergoes proteolytic cleavage,
forming a protein fragment with a molecular mass of 22 kDa. The relative
proportions of these 2 proteins become about equal at 20-30 years of age. Over
time, the protein with molecular mass of 22 kDa predominates in the lens nucleus.
Hejimancik JF, Piatgorsky J. Lens proteins and their molecular biology. In: Abert DM. Takobie
FA, eds. Principles and Practice of Ophthalmology. 3rd ed, Philadelphia: Saunders: 2008:
Val 1, chapter 108.
Increase of Water-Insoluble Proteins With Age
As the lens ages, its proteins aggregate to form very large particles. These particles
become water-insoluble and scatter light, increasing the opacity of the lens. Of
note, the water-insoluble protein fraction increases with age. even if the lens
remains relatively transparent. Conversion of the water-soluble proteins into wat
insoluble proteins appears to be a natural process in lens fiber maturation, but it
may occur more quickly in cataractous lenses.
In cataracts with significant browning of the lens nucleus (brumescent cataracts),
the increase in the amount of water-insoluble protein correlates well with the degree
of opacification. In markedly brunescent cataracts, as much as 90% of the nuclear
proteins may be in the insoluble fraction. Associated oxidative changes occur,
including protein-to-protein and protein-to-glutathione disulfide bond formation
These changes result in decreased levels of the reduced form of glutathione and
increased levels of glutathione disulfide (oxidized glutathione) in the cytoplasm of
the nuclear fiber cells. It is the general view that glutathione is essential to maintain
a reducing environment in the lens cytoplasm. 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. This insoluble protein fraction 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
Carbohydrate Metabolism
The goal of lens metabolism is the maintenance of lens transparency. In the lens,
energy production largely depends on glucose metabolism. Ghicose enters the lens
from the aqueous humor both by simple diffusion and by a mediated transfer
ALo-Crystallins represent about one-third of the lens proteins by mass. In their
native state, they are the largest of the crystallins, with a molecular mass ranging
between 600 and 800 kDa. In addition, they may associate with other crystallins,
yielding complexes greater than 2 « 10°. There are 2 a-crystallin subunits, GA and
GB, each with a molecular mass of approximately 20 kDa, which form heteromeric
complexes containing about 30 subunits. The sequence of the a-crystallins identifies
them as members of the family of small heat-shock proteins, a-Crystallin
complexes bind to partially denatured proteins and prevent them from aggregating.
Their primary function in lens fiber cells appears to be to inhibit the complete
denaturation and insolubilization of the other crystallins.
f.y-Crystallins are subdivided into 2 groups, based on molecular mass and
isoelectric points. The -crystallins, a complex group of oligomers composed of
polypeptides. are encoded by 7 genes. These crystallins have 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. By gel
chromatography. the B-crystallins can be separated into BH (B high-molecular-mass)
and BL (f low-molecular-mass) fractions.
The y-crystallins are the smallest of the crystallins, with a molecular mass in the
range of 20 kDa or less. The native y-crystallins do not associate with each other or
with other proteins and, therefore, have the lowest molecular mass of the crystallin
fractions. In humans, the gamma family is encoded by 4 genes. X-ray
crystallographic studies have determined the 3-dimensional structure of the y-
crystallins fo high resolution. Fourfold repetition of a core 3-dimensional structural
motif suggests that the B.y-crystallins might have arisen from double duplication
and fusion of a gene for a 40-residue polypeptide. The basic structure of the B-
crystallins and y-crystallins has been maintained through hundreds of millions of
years of vertebrate evolution.
Bloemendal H, de Jong W, Jaenicke R. Lubsen NH. Slingsby C, Tardieu A, Aging and vision:
structure, stability and funtion of lens erystalins. Prog Biophys Mo! Biol. 2004:86(3): 407—
485,
Membrane Structural Proteins and Cytoskeletal Proteins
‘The water-insoluble fraction of lens proteins can be further divided into 2 fractions.
1 soluble and 1 insoluble in 8 M urea, The urea-soluble fraction of the young lens
contains cytoskeletal proteins that provide the structural framework of the lens
cells. Microfilaments and microtubules found in lens cells are similar to those found
in other cell types. However, the lens contains 2 types of intermediate filaments
that are unusual: one class is made from the protein vimentin, which is not usually
found in epithelial cells: the other class, the beaded filaments, is composed of the
proteins phakinin and filensin, which are specific to the lens. Genetic disruption of
the structure of the beaded filaments leads to disruption of the structure of the Jens
fiber cells and formation of a cataract.
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