Structure And Functions Of The

Organelles Involved In Protein

Secretion And Modification

Ravinayak Patlavath
Department of Botany,
Faculty of Science,
The M S University of Baroda
Previous lecture recap……..
Protein modification
• The complexity of a protein is increase by adding, removing or
modifying a molecule to the protein.

• As this process occurs after translation, this is known as

‘post-translational modification (PTM) of proteins’

• PTM is required for controlling protein stability, localization,

activity and conformation.
 Types of
Post-Translational Modification
(PTM)
 Fate of a Protein….

1. Transported within the cell (cytoplasm or some organelle)

eg. Enzymes
2. Outside the cell on the plasma membrane eg cell surface
receptors, membrane proteins
3. Outside the cell secreted in the extracellular space
Eg. Insulin secreted by beta-cells of pancreas
Antibodies secreted by plasma cells (used for plasma
therapy for covid19 treatment),
Plant cell wall synthesis enzymes
 Where does PTM occur within the cell?
&
Which are the Organelles involved in protein
secretion and transport?

• Two cell organelles are involved are

1. Endoplasmic reticulum
2. Golgi complex
Endoplasmic reticulum
• Endoplasmic reticulum — ‘a net in the cytoplasm’
 Endoplasmic Reticulum
• Eighteenth-century European ladies carried purses of netting
called reticules.
• The name ‘endoplasmic reticulum’ was first coined by Porter in
1953, who in 1945 had observed it in liver cells under electron
microscope.
Endoplasmic Reticulum
• The ER is the largest organelle in the cell and is a major site of
protein synthesis and transport, protein folding, lipid and
steroid synthesis, carbohydrate metabolism and calcium
storage

• The ER is composed of a continuous membrane system that

includes the nuclear envelope (NE) and the peripheral ER,
defined by flat sheets and branched tubules
Morphology of ER
ER membrane

Nuclear
Peripheral ER
envelope

Sheets

Branched
tubules

Vesicles
Peripheral ER
 Nuclear Envelope
• The nuclear envelope is made up of
two lipid bilayers the inner nuclear
membrane (INM) and outer nuclear
membrane (ONM)
• It shares a common lumen with the
peripheral ER.
• Hundreds of nuclear pores spanning
the ONM and INM of the nuclear
envelope allow transport of
molecules, including RNAs and
proteins, at various rates of diffusion
or regulated transport depending on
the size of the molecule.
 Peripheral ER
• The nuclear envelope is connected to the peripheral ER.
• The Peripheral ER can be morphologically divided into
structures– cisternae, tubules and vesicles.
•The cisternae are •Vesicles are fragments •Tubules forms an
long, flattened, sac of SER or RER interconnecting system of
like, un-branched •These two types of pipelines curving through the
tubules, arranged vesicles have different cytoplasm in which they occur.
parallely in bundles densities, indicating •Tubular form of ER is dynamic
or stacks. different nature of their in nature, i.e., it is associated
• Each sac is separated contents. with membrane movements,
by a cytosolic space. •Vesicles may often fission and fusion between
RER usually exists as remain present in the membranes
cisternae. cytoplasm of most cells •SER is mostly Tubular
 Based on Morphology and Function

ER

Rough ER Smooth ER

RER is bounded by rough walls because ribosomes

remain attached with its outer surface while SER lack
ribosomes hence they appear smooth walled.
Can you identify RER and SER ?
 Structure of Rough ER
• RER is bounded by rough walls because
ribosomes
• The cavity of RER is very narrow and it
contains various enzymes and cofactors
required for PTM
• RER contains two trans-membrane
glycoproteins, such as ribophorin I and
ribophorin II.
• These proteins tend to interact with each
other arid form intramembranous network,
which control the ribosome binding sites in
the plane of the ER membrane.
• RER is highly developed in cells of
pancreas, liver, goblet cells, plasma cells
etc. where active protein synthesis is
involved.
 Structure of Smooth ER
• The smooth ER though forms a
continuous system with the RER,
has a different morphology.
• It’s membrane is smooth and lack
ribosomes.
• In general, SER are found in cells
that are active in glycogen and
lipid metabolism, such as,
adipose cells, interstitial cells,
glycogen storing cells of liver,
spermatocytes and leucocytes
etc.
Which of these two is an active protein secreting cell?
A Plasma cell B Adipocyte
How to observe ER?
• Ribosomes and ER cannot be observes in light microscope. It has to
be observed under electron microscope.
 Where does PTM occur within the cell?
&
Which are the Organelles involved in protein
secretion and transport?

• Two cell organelles are involved are

1. Endoplasmic reticulum
2. Golgi complex
General function of ER in protein
synthesis
Special Functions of RER
• Formation of disulfide bonds.
• Proper folding of proteins
• Addition and processing of carbohydrates/Glycosylation
• Specific proteolytic cleavages
• Assembly into multimeric proteins
• Synthesis of Proteins on Membrane Bound Ribosomes
Formation of disulfide bonds
• The redox environment in the
lumen of the rough ER is favorable
for oxidation of cysteine sulfhydryl
groups (–SH) to disulfide bonds (–
S–S–), whereas this reaction does
not occur in the cytosol. Thus
disulfide bonds are common in
secretory proteins and exoplasmic
domains of membrane proteins,
but are absent from soluble
cytosolic proteins.
• Rearrangement of disulfide bonds
is catalyzed by the enzyme protein
disulfide isomerase (PDI).

https://www.ncbi.nlm.nih.gov/books/NBK21741/
 Proper folding of proteins
• Protein disulfide isomerase
(PDI), an enzyme localized to
the ER lumen, catalyzes the
rearrangement of disulfide
bonds, accelerating the folding
of newly synthesized secretory
and membrane proteins in the
ER.
• The folding of many newly
made proteins within the ER is
facilitated by other folding
catalysts such as peptidyl-
prolyl isomerases and the
lectins calnexin and
calreticulin.
Specific proteolytic cleavages
• Abnormally folded proteins and unassembled subunits are
selectively retained in the ER, either because they form
aggregates or because they are permanently bound to
the chaperone Hsc70 or other ER chaperones.
• Unassembled or misfolded proteins in the ER often are
transported back through the translocon to the cytosol where
they are degraded in the ubiquitin/ proteasome pathway.
• Accumulation of misfolded proteins in the ER lumen induces
increased production of ER protein-folding catalysts via the
unfolded-protein respons.
• Certain resident ER proteins are retained in the ER by a C-
terminal KDEL sequence or are retrieved there from the cis-
Golgi network by the KDEL receptor.
Assembly into multimeric proteins
• Assembly of subunits to
form multimeric secretory
and membrane proteins occurs in the ER.
• Only properly folded proteins are
transported from the rough ER to
the Golgi complex of vesicles.
• Immunoglobulins, which contain two
heavy (H) and two light (L) chains, all
linked by S–S bonds.
• Hemagglutinin (HA), the trimeric protein
that forms the spikes protruding from the
surface of the influenza virus particle

https://www.sigmaaldrich.com/technical-documents/articles/biology/antibody-basics.html
Addition and processing of
carbohydrates/Glycosylation
• Many of the proteins that enter the ER lumen or ER membrane are converted to glycoproteins
by the action of oligosacchayl transferase (active site present on the luminal side of ER
membrane).
• Oligosaccharide composed of N-acetylglucosamine, mannose, and glucose, and containing a
total of 14 sugars.
• oligosacchayl transferase is a membrane bound enzyme catalyzes the covalent transfer of
oligosaccharide attached to a specialized lipid called dolichol in the ER membrane to the
amino (NH2) group of an asparagine side chain on the protein.
• The oligosaccharides on proteins/Glycoprotein can serve various functions. Like protect a
protein from degradation, hold it in the ER until it is properly folded, or help guide it to the
appropriate organelle by serving as a transport signal for packaging the protein into
appropriate transport vesicles.
• Oligosaccharide when displayed on the cell surface form part of the cell’s outer carbohydrate
layer or glycocalyx and can function in the recognition of one cell by another.
• Asparagines that are glycosylated are always present in the tripeptide sequences asparagine-
X-serine(Asp-X-Ser) or asparagine-X-threonine(Asp-X-Thr), where X can be almost any amino
acid.
https://www.researchgate.net/figure/N-linked-and-O-linked-protein-glycosylation-occurs-in-the-ER-and-Golgi-apparatus-The_fig3_258445421
Home work
Special Functions of Smooth ER
1. SER helps in detoxification of a wide variety of xenobiotics
(toxic materials of both endogenous and exogenous origin) in
the liver such as, barbiturates, ethanol, aspirin and petroleum
products.
2. Glycogenolysis and Blood Glucose Homeostasis
3. The regulated release of Ca++ from SER membranes triggers
specific cellular responses including, the contraction of
skeletal muscle cells.
4. SER have a key role in the synthesis of cholesterol, the
precursor of steroid hormones and bile acids.
5. Synthesis of Lipids.
Detoxification of Drugs
• Detoxification is the process by which toxic substances
are converted into more soluble & less toxic substances,
which are mainly eliminated through urine & bile.
• Takes place in the “liver” and Kidney.
• The compounds to be detoxified include Drugs –
antibiotics, cardiac drugs, steroids, etc.
• Enzyme involved is cytochrome P450 (Monooxygenase or
mixed function oxidases)
• The most common reaction catalyzed by cytochrome P450
is a monooxygenase reaction (Oxidation),
• P450 family of proteins prevalent in SER of hepatocytes
also found in lung and intestinal cells.
• Electron transport system of ER transfers e- stepwise
from NADPH or NADH to cytochrome p450.
• P450 gets reduced and it donates e- to molecular O2 .
Molecular O2 gets activated for hydroxylation.
• NADPH-serves as e- donor for drug detoxification and
sterioid biosynthesis.
• NADH-serves as e- donor during hydroxylation of
fattyacids.
NADPH + H+ + O2 + RH  NADP+ + H2O + R-OH
Glycogenolysis and Blood Glucose
Homeostasis
• Glucose-6-
phosphatase is a
membrane bound
enzyme found in
liver, kidney and
intestine SER.
• Brain and muscle
directly use up
Glucose-6-
phosphate.

http://www.mun.ca/biology/desmid/brian/BIOL2060/BIOL2060-12/12_03.jpg
The regulated release of Ca++ from
SER membranes

https://www.researchgate.net/figure/Cardiac-muscle-cell-contains-both-sarcoplasmic-reticulum-SR-and-endoplasmic-reticulum_fig1_42805931/download
• The Sarcoplasmic Reticulum is responsible for excitation-
contraction coupling of cardiomyocytes.
• For contraction, Ca 2+ is released from the SR through the
ryanodine receptor (RyR) Ca 2+ channel and then Ca 2+ is taken up
by the sarcoplasmic/endoplasmic reticulum Ca 2+ ATPase (SERCA)
of the longitudinal SR domains causing relaxation.
• Cardiac muscle cells may also contain ER. Ca 2+ fluxes from the ER
are required for cardiac development and housekeeping functions
of cardiomyocytes.
• In the lumen of the ER, Ca 2+ is stored bound to Ca 2+ binding
proteins including calreticulin (CRT), protein disulfide isomerase
(PDI), Glucose regulated protein 94 (Grp94), Immunoglobulin
binding protein (BiP), and ERp57. These proteins play a critical role
in folding and posttranslational modification of newly synthesized
proteins.
SER is involved in synthesis of Lipids
• It synthesizes lipids, phospholipids, and steroids. Cells
which secrete these products, such as those in
the testes, ovaries, and sebaceous glands have an
abundance of smooth endoplasmic reticulum.
• Phospholipids, ceramide, and sterols are primarily
synthesized in mammalian cells by enzymes in the ER,
usually associated with the cytoplasmic leaflet of the SER
Summary