Lysosomes are found in all animal cells, but are most numerous in disease-fighting cells, such as white blood

cells. This is because white blood cells must digest more material than most other types of cells in their quest to battle bacteria, viruses, ana other foreign intruders. Several human diseases are caused by lysosome enzyme disorders that interfere with cellular digestion. Tay-Sachs disease, for example, is caused by a genetic delect that prevents the formation of an essential enzyme that breaks down complex lipids called gangliosides. An accumulation of these lipids damages the nervous system, causes mental retardation, ana death in early cnildhood. Also, arthritis inflammation and pain are related to the escape of lysosome enzymes.

Occurence
Lysosomes are found in most of the animal cells and few plant eells.The hepatic cell, pancreatic cells, kidney cells, etc. posses large number of lysosomes. According to Matile (1969) the lyososome in plant cell are membrane bound. Generally lysosome are absent in the prokaryotic cells. Slime molds also have lysosome. Lysosome are pleomorphic as seen in the meristematic cells of root. The diameter may be 0.4U to o.8u ;but in mammalian kidney cell lysosomes as long as 5 u have been found.

Formation of Lvsosomes
The hydrolytic enzymes are formed in the RER. Enzymes have a terminal mannose-6-phosphate group as a marker to be packaged in lysosomes. They are packaged at the trans face of the Gogi complex (termed the Trans Golgi Network -TGN). The newly formed vesicles that bud off are termed early endosomes or primary lysosomes. They are small and have homogeneous, electron-dense contents.

re
S Lysosome are saccate structure filled with digestive enzyme. Each lysosome has two parts, vizjimiting membrane and inner dence mass.

i)Limitting membrane:
Sit is not double bound membraned like in other organelles. It is single and composed of lipoproteins. The membrane however is bilayered like any unit u membrane. 325S,i*

P R O T E I NS / E NZ Y M E S

2)Inner dence mass:

f i t is the core of lysosome; it mostly enzymatic.The membrane is impermeable to substrate the enzyme of which are present in the lysosome. S Ocassionaly certain substance called labilizers detabilize the membrane causing the enzyme to come out. Alternatively, there are substance called stabilizer which stabilize the membrene. ^The limited permeability of the mambrane prevent excessive digestion of cellular component by the lytic enzyme of lysosome. v"The enzyme present in the lysosome are-Nuclease,Phosphatase,Lipage,Glycosidase and Sulphatases.

> Lysosomes are polymorphic assuming various shapes depending on the cell type and situation. Basicallly there are two types of lysosome Viz, primary lysosome 2) secondery lysosome 1) a) Phagosomes b) Residual bodies c)Autohagic vacuoles 1) Primary lysosome There are sac like stucture produce like directly fro ER from the golgi bodies.
2) secondery

lysosome

a) Phagosomes:These are also known as heterophagosomes or digestive vacuoles. When a foreign gene enters the cytoplasm, the plasma membrane forms asac around these substance. These are around these

substances. These are the phagosome or pinosomes.The lytic enzyme present in phagosome digest the foreign substance. The phenomenone is called Phagocytosis or pinocytosis.

b) Residual bodies:After the digested meterial has pas out of the lysosome, the undigested material remains within the lysosome. Such lysosome with residual substrate are called residualbodies.lt may remain within the cell or very often they are pushed out of the cell by exocytosis. c)Autophagic vacuoles :Ocassionally, a lysosome may digest a part of the cell(ER,mitochondria).This is a mechanism to digest the a part of cell without destroying the entire cell.Iiver cells show autophagic vacuoles or autophagosome under starvation.

Origin of lysosome

a)Extra cellular origin : Vacuoles absorbed due to pinocytosis or phagocytosis may become secondery lysosome after fusing with primary lysosome. b)Origin from golgi bodies: There are enough evidence to that lysosome are produce from golgi bodies at the height of their secretory activity. c)Origin from ER : Acorrding to Novikoff (1965), the rogh ER gives rise to the lysosome.

COURSE NO: GP 508 (2+1)

CELL BIOLOGY AND MOLECULAR GENETICS

three special properties

i)The Proton pump 2)Gtyco protein coat ^Transporter channel

i) The proton pump:

S It an ATP driven proton [H+] pump to maintain a low pH (4.5-5.5) in the lysosomal compartment.

Has a lollipop' structure similar to the F1 and F of mitochondria, and consisting of.
S A head containing 6 polypeptide units.

s A stalk contains 5 polypeptide units.

S It is inhibited by n-ethylmaleimide.

2) A glycoprotein coat:
V rich in carbohydrates, on its inner surface to protect it against hydrolysis by its own enzymes

 Transporter channels that transport break-down products such as amino acids, glucose, nucleotides and other small molecules out of the lysosome. These molecules may move out,
S by facilitated diffusion; S by

active transport; S by co-transport using the proton (H+) gradient to provide the energy for transport. S They early endosomes (primary lysosomes) may fuse with phagocytic or other vesicle to form late endosomes or secondary lysosomes.

Major function oflysosome:

The main function of lysosome is digestion of intra celluler substances. Acorrding to Hirsch and Coh (1964), lysosome help in destyoing the dead cell from tissue. Recently it has been found out that in some animals (frog), lysosome play an important role in metamorphosis. Lysosome also perhaps help in the formation and destruction of bone cells. They digest ingested material and aged or damaged organelles

Functions of Lysosomes

Lysosomes may be involved in following pathways:

1.
2.

Phagocytosis Autophagocytosis

3. Formation of endosomes

4. Receptor-mediated endocytosis

osis:
Phagocytosis is the process by which the cell engulfs particulate matter (>o-5 m) from the extra cellular space, and digests it by lysosomal action. Three main cell types are capable of performing this function.

a) Macrophages:
are widely distributed in the connective tissue around the body phagocytose and digest all particulate matter in the extracellular space, including micro-organisms, foreign particles and damaged cells. are derived from monocytes, circulating in the peripheral blood. may have different names in different tissues such as Kupffer cells in the liver, osteoclasts in bone, and microglia in the central nervous system. b) Neutrophils: In the peripheral blood, which phagocytose and digest microorganisms. c) Eosinophils: In the peripheral blood, which phagocytose and digest antigen-antibody complexes.

Phagocytosis
Adhesion of the particle to the glycxjcalyx or specific receptor on the plasma membrane . Extrusion of pseudopodia to surround the particle. This is mediated by actin filaments. . Formation of a phagocytic vacuole containing the engulfed particle - Fusion with a primary lysosome

Binding

Phagocytosis

Endocytotic vesicle

Secondary lysosome

Residual body

2) Autophagocytosis:

This is the process in which old or damaged organelles are broken down. It occurs in practically all cells as a recycling system. It is most marked in cells that are not replaced, such as neurons.

Autophagocytosis
i. The organelle is surrounded

Organelle

by vesicles, which coalesceThe coalesced vesicles form a membrane surrounding the organelle. This is called an autophagic vacuole. 3. The autophagic vacuole fuses with one or more primary lysosomes to form secon lysosomes. 4. Residual bodies are lysosomes with partially undigested material.
2,.

Autophagic vacuole

Secondary lysosome

Residual body

Dr. S.R.Patel Associate Prof.; Dept. of botany; N.A.U,Navsari.

Movaliya Darshan j. First sem. M.sc(agrL) Dept. of Botany; N.M. CoUege of Agriculture,Navsari

osis and exocytosis Endocytosis is the uptake of extra cellular fluid by infolding of the plasma membrane, and formation of a vesicle containing the extra cellular material. This process was formerly called pinocytosis. Exocytosis is the reverse of pinocytosis, i.e. the extrusion of fluid contained in vesicles into the extra cellular space. Endocytosis and exocytosis are important mostly for membrane flow. For example, exocytosis replaces the plasma membrane removed by phagocytosis. Similarly, exocytosis of secretion vesicles must be balanced by endocytosis.

4) Receptor-mediated endocytosis: Receptor-mediated endocytosis is the process whereby cells that have a specific receptor take up specific macromolecules. This process is used for the uptake of hormones, growth factors, antibodies, lipoproteins etc. The receptors are integral membrane proteins. The molecule that binds to the receptor is termed the ligand.

Receptor-mediated
endocytosis
Binding of the ligand to the receptor. Accumulation of clathrin, adaptor protein and dynamin on the cytoplasmic surface of the plasma membrane at a particular site. Formation of a pit, and accumulation of the ligand receptor complex at the site of clathrin acculmulation. Deepening of the pit clathrin-coated vesicles ligand-receptor complex. and formation of containing the

jl

-z- Lateral diffusion of the ligand-receptor complex.

3.

4.

'B-

6. The vesicles lose their clathrin coat 7. The vesicles fuse with a primary lysosome (early endosome) and the ligand is cleaved from the receptor.

8.

Familial hypercholesterolaemia: is a condition in which there is defective binding of low density lipoprotein (LDL) to its receptor. Receptor-mediated endocytosis of the LDL does not occur, and it accumulates as high levels in the blood.

 Peroxisome are found in animal cell and also in the leaves of higher plant. These particles participate in the oxidation of substrate resulting in the formation of hydrogen peroxide. In plants, peroxisome are involved in photorespiration. Structurally peroxisome are variable in shape and size but are gnerally circular. They have an envalope of a single membrane made up of protein and lipids enclosing a granular metxix. In certain instances, the metrix is crystaline or amorphous. The peroxisome catalize the H202 production. In plants, the peroxisomes contain enzymes of glycolate patheway.

PEROXISOMES AS SEEN IN A
HUMAN LIVER SECTION

History of Peroxisomes
* First observed by electron microscopy in animal cells (1950s), then in plant cells (1960s).

* Christian deDuve (1965) Isolated from liver cells by centrifugation 9 Called them peroxisomes because they generate and destroy H2Oz

Thel e7oxisome
5

Single membrane V Roughly spherical

* 0.2 - 1.7JU11

V Composition varies

Number and Size of Peroxisomes Vary Depending on Environment

o M o d e l s f o r P e r o x i s o

m e B i o g e n e s i s
Subset 2 ot membrane proteins Preperoxisomal template

_____ 1________

Subset 2' of membrane proteins Subset 1 of membrane proteins

E a

_____ 1________
r l y p e r o x i s o m a l p r e c

Nascent peroxisomes

u r s o r s

memb'ane proteins Prepe^ox somal Subset 2 ol endo^eribrane membrane proteins Subset 1 of

__ 1____

__ I ____
Nascent peroxisome

I arly peroxisomal precursor N a t u r e R e v i e w s | M o l e c u l a r C e l l B i o l o g y

Metabolic Functions of^*^ Peroxisomes

Yeasts
Biosynthesis: lysine @Dearadation: amino acids, methanol, p-oxidation of fatty acids, decomposition of hydrogen peroxide, glyoxylate cycle.

Fungi

@> Biosynthesis: penicillin Degradation: p -oxidation of fatty acids, decomposition of hydrogen peroxide, glyoxylate cycle.

Microbodies Lysosomes Peroxisome Glyoxisome

@ Degradation: purines, some reactions of photorespiration (the

conversion of glycolate to glycine and of serine to glycerate), p-oxidation of fatty acids, decomposition of hydrogen peroxide, glyoxylate cycle.

Mammals @ Biosynthesis:

ether phospholipids (plasma log ens), cholesterol and bile acids, polyunsaturated fatty acids a-oxidation of fatty acids, p-oxidation of fatty acids, decomposition of hydrogen peroxide.

@ Degradation: amino acids, purines, prostaglandin, polyamines,

Humans @ Biosynthesis:

ether phospholipids (plasma log ens), cholesterol and bile acids, polyunsaturated fatty acids. p-oxidation of fatty acids, decomposition of hydrogen peroxide.

@ Degradation: amino acids, purines, a-oxidation of fatty acids,

Peroxisome in Plants
9 9 9 9 9 P -oxidation of fatty acids

Glyoxylate cycle Photorespiration (Glycolate pathway) Degradation of purines Decomposition of hydrogen peroxide

wo Types of Peroxisomes in Plants

9 Leaves * Catalyzes oxidation of side product of C02 fixation in photorespiration. 9 Germinating seeds * Converts fatty acid in seed lipids into sugars needed for growth in the young plant.

The Glycolate Cycle

CHLOROPLAST
R ib u l o s e 1 , 5 - b is p h o s p h a te
C02

G lu ta m in e

- P h o s p h o g ly c e r a te

^cr
H20

-Ct

.po --ooc^
3 2

Glutamine Synthetase
G lu ta m a te

P h o s p h o g ly c o la te

P.
PEROXISOME

MITOCHONDRION
oc
C
2

GlycoUcacia ^
j
oxidase

G ly c o la te

2O2 *

I
H202 production
G ly o x y la te
CO

NH.

2 G ly c in e
NH 3 S e r in e

Purine Degradation
Nucleic acid purine moieties (adenine and guanine) are degraded to uric acid

xanthine

o2 h2o2 . uric acid

allantoin

L,02

Xanthine oxidase

Urate oxidase

 High urate oxidase concentrations contribute to formation of crystalline inclusions All purine degradation leads to uric acid

hypoxanthine

xanthine oxidase

xanthine

xanthine oxidase

uric acid

Oxidases
The oxidases use molecular oxygen to remove hydrogen atoms from specific organic substrates A variety of compounds, including L-amino acids, D-amino acids, polyamines, methanol, urate, xanthine, and very-long-chain fatty acids, serve as substrates for the different oxidases

Peroxide Detoxification
9

Oxidases use 02 to oxidize organic substances and produce hydrogen peroxide (H202) e.g., H202 generated by glycolate oxidase reaction, P-oxidation of fatty acids. Peroxisomes also contain catalase, the enzyme that degrades H202.

Importance of H202 degradation

2H202
atdase

2II20+02

Peroxisomes contain a high concentration of catalase, a heme protein

H--O--O--H HO-OH (?)

Other reactive oxygen species (ROS) are formed in peroxisomes

Reactive Oxygen Species

ler
m

le"

le" H202 ^ OH-+ OH h*p H20

le"

.0-0' 'O'O:

H:0:0:H H:0:+ O^H H:0-H

Hydrogen peroxide Hydroxyl radical

m m

m m

Superoxide anion (radical)

# Cause damage to lipids, proteins, DNA 4 Amount ROS is reduced by catalase, and superoxide dismutase (SOD) 202" ------------- - 02 + H202

Microbodies:
The cells of plants, fungi, certain protozoa and liver cells of vertebrates contain certain small granular structures associated with ER, mitochondria and chloropasts. These spherical bodies ranging in size from o.2\i to 1.5a have a central cristalline core and are called microbodies.

Peroxisoma
Kvy _4jGlulamntr:glyoxylrtir nmliMHniissroi.iv IransfonWi*

zymes

J | RUHIIOSC-I.S-hlsphnsphatf rariK>xvtas4>/oxywna<i*' 2 \ PhosphoRlycolate phosplutasc 3 1 Clytolate oxidase S | <;1><-IIMT ttt^rtHHixylrtN.' .: I serin*- hydmxyiiwlliyl

6 \ SrrtiH:RlyoxylflU' amlnotransffcraM

_7J I lydroxypyruv.U- rt-ducl.u*-SJClyccraip kinase 9j Caialase

Glyoxisome
*Glyoxisome are found in the cells of yeast and certain other fungi(Neurospora) and also seeds (oil reach) of many higher plant. +They are similar to peroxisome except that theire core consists of dence rods of 6.o|im in diameter.
* Functionally the glyoxisome contain enzymes for fatty acids mtabolisome, involved in the conversion of lipids to carbohydrate during germination. The glyoxisome play an important role in the glyoxisome cycle. The enzyme Urate oxidase and allantoinase are also found in glyoxisomes.

+ Urate oxidase converts uric acids to allantonin,while allantonase hydrates allantonin to allantonic acid during the degrading of purines.

Lysosomes
(Gr. Lyso=digestive, soma=body) First discovered in 1949 as dence round bodies ,lysosomes were called pericanalicular bodies. It was de Duve,(i955) who gave the present name. They are membrane bound organelles. They are contain hydrolytic enzymes.They have acidic contents (pH 4.5-5.5) and electron-dense heterogeneous contents.

 The discovery of lysosomes involved the use of a centrifuge to separate the various components of cells. In the mid-twentieth century, the Belgian scientist Christian Reni de Duve was investigating carbohydrate metabolism of liver cells and observed that that the cells released an enzyme called acid phosphatase in larger amounts when they received proportionally greater damage in the centrifuge. To explain this phenomenon, de Duve suggested that the digestive enzyme was encased in some sort of membrane-bound organelle within the cell, which he dubbed the lysosome. After estimating the probable size of the lysosome, he was able to identify the organelle in images produced with an electron microscope.

Anatomy of the Lysosome

Plasma Membran e

Lipid Bilaye r

Figure

Hydrolyti c nzvm hnzyi e Mixture

Glycosylated Membrane Transport Proteins

Lysosomes are spherical organelles contained by a single layer membrane, though their size and shape varies to some extent. This membrane protects the rest of the cell from the harsh digestive enzymes contained in the lysosomes, which would otherwise cause significant damage. The cell is further safeguarded from exposure to the biochemical catalysts present in lysosomes by their dependency on an acidic environment. With an average pH of about 4.8, the lysosomal matrix is favorable for enzymatic activity, but the neutral environment of the cytosol renders most of the digestive enzymes inoperative, so even if a lysosome is ruptured, the cell as a whole may remain uninjured. The acidity of the lysosome is maintained with the help of hydrogen ion pumps, and the organelle avoids self-digestion by glucosylation of inner membrane proteins to prevent their degradation.