Lysosomes and Peroxisomes

M.Nagalingam
PhD Scholar (P-1737)
IVRI, Izatnagar

Introduction
Lysosomes (Gr., lyso=digestive + soma=body)
Lysosomes are dynamic organelles that receive and degrade
macromolecules from the secretory, endocytic, autophagic and
phagocytic membrane-trafficking pathways.
The lysosomal lumen is maintained at an acidic pH (around 5) by
an ATP-driven proton pump in the membrane.
C. de Duve and his coworkers (1963, 1964, 1974) worked in
Belgium and their approach was biochemical one.

Alex Novikoffand his research group (1962, 1964) worked in

United States and their approach was morphological and
cytochemical
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Lysosomes
The lysosomes occur in most animal (except mature
erythrocytes) cells
The lysosomes are round vacuolar structures which remain
filled with dense material and are bounded by single unit
membrane.
Their shape and density vary greatly.
Lysosomes are 0.2 to 0.5m in size. Since, size and shape of
lysosomes vary from cell to cell and time to time (i.e.they are
polymorphic), their identification becomes difficult.

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Classes of proteins :
Soluble lysosomal hydrolases (also referred to as acid
hydrolases) 50 known lysosomal hydrolases
Integral lysosomal membrane proteins (LMPs)
~25 LMPs
lysosome-associated membrane protein 1 (LAMP1)
LAMP2,
lysosome integral membrane protein 2 (LIMP2; also
known as SCARB2)
tetraspanin CD63
Adaptor proteins AP1, AP3
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Trans Golgi Network

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Targeting of lysosomal proteins by phosphorylation of

mannose residues

Phosphate groups to the 6 position of mannose residues

N-acetylglucosamine phosphates to lysosomal proteins from UDP-Nacetylglucosamine
Recognition by signal patches
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Endocytosis and lysosome formation

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Possible interaction between endocytosis and biogenesis

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Early endosomes
early endosomal antigen 1 (EEA1) and Rab5 are widely
used as markers

Late endosomes vs lysosomes

lack of M6P receptors in lysosomes

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Clathrins

Recycling of M6PRs by endosome-to-TGN carriers will not require clathrin ,

but requires the retromer subunit sorting nexin 1 (SNX1) and/or SNX2
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Mannose 6 phosphate independent pathway

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Degradative function

Two major degradation routes:

The lysosomal network and
the ubiquitin-proteasome system .
Proteases
Cathepsin family of proteases
serine (A and G),
Cysteine (B, C, F, H, K, L, O, S, V, W, and X), and aspartic
cathepsins (D and E).
pH: Vacuolar H+-ATPase, a transmembrane multimeric protein
complex

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Lysosomes in phagocytosis and autophagy

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Autophagy in mammalian cells

Chaperone-mediated autophagy,
Microautophagy,
Macroautophagy
mammalian target of rapamycin (mTOR)

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Lysosomal exocytosis
Lysosomal exocytosis plays a major role in important processes
such as immune responses, bone resorption, cell signaling, and
plasma membrane repair

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Evasion of lysosome fusion by microbes

Preventing lysosome fusion - Escherichia coli K1
Delaying phagolysosome biogenesis- Salmonella
enterica and Mycobacterium tuberculosis
Escaping the phagosome-Listeria, Shigella and
Rickettsia
Legionella pneumophila, Coxiellabrunetti and Brucella
abortus, can reside in an autophagosomal compartment
where they multiply

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Lysosomal storage disorders

Over50 human lysosomal storage conditions have been recognized, and

although individually rare, their combined prevalence is1in 8000 births
Most of these disorders are autosomal recessively inherited such as NiemannPick disease, type C, however a few are X-linked recessively inherited, such as
Fabry disease and Hunter syndrome (MPS II).
Lysosomal storage disorders frequently involve the central nervous system

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Lysosomal storage disorders

Tay-Sachs disease was the first lysosomal storage

disorder (LSD) described, in 1881
Ernest GAUCHER
(1854-1919)

Gaucher disease was the second, in 1882

The first link between an enzyme deficiency and a
LSD (-glucosidase and Pompe disease) was
published in 1963 by Hers

The successful treatment of a LSD, Gaucher

disease with -glucosidase, became available in the
early 1990s
Pompe became the first disease formally
recognized as a lysosomal storage disorder.

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Biochemical and Cellular basis of LSDs

1 catalytic activity
2 activator
3 misfolding
4 multienzyme complex
5 glycosylation

6 M-6-P targetting
7 other transport steps
8 membrane transporters
9 membrane regulators

Futerman AH & van Meer G (2004) 5:554-565

Lysosomal storage disorders

Type of defect protein

Disease examples

Deficient protein

Lysosomal enzymes primarily

Tay-Sachs disease, I-cell

disease,Sphingolipidoses
(e.g., gangliosidosis, Gaucher
and Niemann-Pick disease)

Various

Posttranslational modification of
enzymes

Multiple sulfatase deficiency

Multiple sulfatases

Membrane transport proteins

Mucolipidosis type II and IIIA

N-acetylglucosamine-1-phosphate
transferase

Enzyme protecting proteins

Galactosialidosis

Cathepsin A

Soluble nonenzymatic proteins

GM2-AP deficiency, variant

AB, Niemann-Pick disease, type C2

GM2-AP, NPC2

SAP deficiency

Sphingolipid activator proteins

Niemann-Pick disease, type C1

NPC1

Salla disease

Sialin

Transmembrane proteins

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I - Defective metabolism of glycosaminoglycans

" the mucopolysaccharidoses"
MPS I

(Hurler, Hurler-Scheie, Scheie)

MPS II (Hunter)
MPS III (San filipo Types A,B,C and D)
MPS IV (Morquio type A and B)
MPS VI (Maroteaux-Lamy)
MPS VII (Sly)
MPS IX (Hyaluronidase deficiency)
Multiple Sulfatase deficiency

II - Defective degradation of glycan

portion of glycoproteins
Aspartylglucosaminuria
Fucosidosis, type I and II
Mannosidosis
Sialidosis, type I and II

III - Defective degradation of glycogen

Pompe disease

IV - Defective degradation of sphingolipid

components
Acid sphingomyelinase deficiency (Niemann-Pick A & B)

Fabry disease
Farber disease
Gaucher disease, type I, II and III

GM1 gangliosidosis, type I, II and III

GM2 gangliosidosis (Tay-Sachs type I, II, III and Sandhoff
Krabbe disease

Metachromatic leukodystrophy, type I, II and III

V - Defective degradation of polypeptides

Pycnodysostosis

VI - Defective degradation or transport of

cholesterol, cholesterol esters, or other
complex lipids
Neuronal ceroid lipofuscinosis, type I, II, III and IV

I-cell disease
Caused by the deficiency of an enzyme(N-acetyglucosamine
phosphotransferase) Because of the absence of this enzyme,the
secreted enzymes lack the mannose phosphate residues which
is a signal for targeting lysosomal enzymes to lysosomes.
Symptoms:a rare inherited metabolic disorder characterized by
coarse facial features,skeletal abnormalities and mental
retardation .Many cells from these patients contain lysosomes
that are bloated with undegraded materials.
back

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Peroxisomes

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Peroxisomes

C.de Duve and P. Baudhuin (1966) coined the term peroxisome for the
micro-bodies of mammalian systems and studied their structure and
function.
Called them peroxisomes because they generate and destroy H2O2
All animal cells (except erythrocytes) contain peroxisomes.
Peroxisomes are related to specialized peroxisomes called glycosomes in
parasites such as Trypanosomes, and to plant glyoxysomes

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~60 known enzymes in the matrix and ~45 documented

integral or peripheral membrane proteins
Peroxisomes are surrounded by a single membrane and they
range in diameter from 0.1 to 1m
They have crystalline and non-crystalline inclusions.

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Biogenesis
(a) peroxisome proliferation by division,
(b) peroxisome de novo biogenesis,

Dynamin-related proteins (DRPs), and Pex11-type

peroxisome proliferators (PPPs), strongly involved in
controlling peroxisome number and division
Most of proteins that reside in the peroxisome matrix and
membrane are synthesized in the cytosol and then imported
post translationally to the organelle.
About 25 PEX genes, encoding proteins named peroxins,
are necessary for the biogenesis of the organelle.

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Assembly of peroxisomes

Pex3 is an integral
transmembrane
protein
Pexl9
is
a
farnesylated protein
found largely in the
cytosol.

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Protein Import
C-terminal signal
sequence: SKL
(PTS1)
N-terminal signal
sequence: RLX5HL
(PTS2)
Proteins involved in
import: peroxins
Import driven by
ATP hydrolysis
Dont have to be
unfolded for import

Functions
Hydrogen peroxide metabolism
Enzymes involved in the degradative oxidation (e.g., -oxidation of very
long chain fatty acids, 2-methyl-branched fatty acids, dicarboxylic acids,
leukotrienes, bile acid intermediates and cholesterol side chains, and both
a-andb-oxidation of 3-methyl-branched chain fatty acids);
The early steps in the synthesis of ether glycero-lipids or plasmalogens;
The formation of bile acids, dolichol, and cholesterol; and
The catabolism of purines, polyamines, and amino acids, and the
detoxification of reactive oxygen species such as hydrogen peroxide,
superoxide anions, and epoxides. In methylotrophic yeasts, peroxisomes
are also involved in the metabolism of methanol and methyl amines.

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peroxisomal diseases
Genes mutation
proteins
involved
in the
uptaking

Empty peroxisomes

Machinery
for
transport

ZS

VLCFAs accumulate
in the brain

A single peroxisomal
enzyme absence
Defect in a
membrane protein
that transports VLCFAs
ALD : adrenoleukodystrophy

Enzymes
fail to
be
imported

ALD

ZS: Zellweger syndrome

VLCFAs: Very-long-chain fatty acids

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Thank you

Acknowledgement

Literature from internet

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PEROXISOMAL MATRIX PROTEIN IMPORT

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