Membrane compartments of the

cell
Cytoskeleton

Svetlana Lutsenko, Dept. Biochemistry and Mol. Bology

MRB 624, Ext. 4-6953, lutsenko@ohsu.edu
Reading for the “Cytoskeleton” portion:

Alberts “Essential Cell Biology” pp.513-532

Mechanisms by which molecules can pass through membranes

1. Transport of various molecules,

including proteins, DNA and RNA
through large protein-lined pores.
Specificity of transport is based on
size exclusion

3. Simple diffusion and protein-

mediated transport of ions and small
molecular weight nutrients, such an
amino-acids and sugars

5. Vesicular transport is mediated by

membranes that bud off as a vesicles
from their original compartment and
carry out specific cargo (ions,
proteins, neurotransmitters) to
another compartment
Cellular Compartments and Their
Functions
• Cytosol : many biosynthetic pathway;
synthesis of cytosolic protein
• Nucleus : DNA and RNA synthesis
• Mitochondria : energy metabolism,
urea cycle
• Endoplasmic Reticulum: Protein
Secretion; Synthesis of Membrane
Proteins
• Golgi : Distribution and Modification of
Secreted Proteins
• Lysosomes : Degradation of proteins
and other molecules
• Peroxisomes : Degradation of Certain
Fatty Acid
 Nucleus
• The nucleus is a membrane
bound structure that
contains the cell's hereditary
information and controls the
cell's growth and
reproduction.
• It is commonly the most
prominent organelle in the
http://biology.about.com/library/weekly/aa032300a.htm
cell
Nuclear Membranes
• The nuclear content is located in the
nuclear lumen and is surrounded by a
double membrane or nuclear envelope,
envelope
composed of inner membrane and outer
membrane.
membrane

• The outer membrane is contiguous with

the ER

• The nuclear membrane contains nuclear

pores,
pores which provide selective access into
and out of the nuclear lumen

• The inner membrane has a protein lining

called the nuclear lamina, which binds to
chromatin and other nuclear components.
 The Nuclear Pore Complexes
form a continuous aqueous channel between cytoplasm & nucleoplasm

nuclear pore complex is

large –about 120 million
Daltons

-30 different proteins found in pore; basic subunit

repeated 16 times. “Central granule" now called
"transporter" functions to move molecules through
pore. There is a ring of proteins that anchors pore
to N.E. and the "basket" of fibers with unknown
function
-no obvious motor proteins were found

Proposed mechanism -transport factors with cargo destined to pass through nuclear pore bind to
pore increasing their local concentration. Higher local concentration allows diffusion across the
diffusion barrier channel
 The Nucleolus
• The region of the nucleus where portions of chromosomes that contain genes
coding for ribosomal RNA are transcribed and ribosomal subunits are
assembled
•Stretch of DNA with rRNA genes
nucleolar organizing region =
(NOR)
• Ribosomal proteins are synthesized
in cytoplasm and transported into
the nucleus
•These proteins self associate with
appropriate rRNA during rRNA
synthesis forming immature
http://biology.about.com/library/weekly/aa032300a.htm ribosomal subunits
•Ribosomes finish self assembly in
cytoplasm
Endoplasmic Reticulum
 The Endoplasmin Reticulum (ER) is an extensive,
extra-nuclear membrane system with the following
functions:

• ER is a home for various enzymes involved in protein folding , drug

detoxification, membrane lipid biosynthesis, cholesterol and fatty acid
metabolism

• ER is an entry point for protein sorting. Targeting of these proteins is

mediated by signal sequence. The membrane proteins are inserted into
the ER membrane in their proper orientation. Secreted proteins are
translocated into ER lumen and then transported to the destination
place
 Rough and Smooth ER
• Two regions of the ER differ in both structure and function. Rough ER has ribosomes attached
to the cytoplasmic side of the membrane. Smooth ER lacks attached ribosomes. Typically, the
smooth ER is a tubule network and the rough ER is a series of flattened sacs.

The smooth ER has a wide range of

functions including carbohydrate and
lipid synthesis.

It serves as a transitional area for

vesicles that transport ER products to
various destinations.

In liver cells the smooth ER produces

http://esg-www.mit.edu:8001/esgbio/cb/org/er.gif
enzymes that help to detoxify certain
compounds. In muscles the smooth ER
assists in the contraction of muscle cells
and in brain cells it synthesizes male
and female hormones.
The rough ER manufactures membranes and secretory proteins.
In leukocytes the rough ER produces antibodies.
In pancreatic cells the rough ER produces insulin.

The rough and smooth ER are usually

interconnected and the proteins and
membranes made by the rough ER move
into the smooth ER to be transferred to
other locations.

The cytoplasm has a reducing environment,

while ER lumen is oxidizing . This
difference is generated by unequal
distribution of trypeptide glutathione and
is essential for formation of disulfide bonds
in proteins and for proper folding
Mitochondria
 Mitochondria (singular: mitochondrion) are the sites of
aerobic respiration, and generally are the major energy
production center in eukaryotes

The number of mitochondria range from one to thousands per cells.

They are often positioned in cells nearest to sites of energy utilization

One of the richest sources of mitochondria is a hummingbird flight
muscle
 Mitochondria are a double membrane organelle in which the
inner membrane is in-folded to form “cristae”.

The outer membrane is a fairly

simple phospholipid bilayer,
containing porins, proteins that
render it permeable to molecules
of about 10 kilodaltons or less.
Ions, nutrient molecules, ATP,
ADP, etc. easily pass through the
outer membrane and enter the
intermembrane space
The inner membrane is more
complex and contains respiratory
chains and transporters

The matrix lies within the inner membrane. The access to this compartment
often requires specific transporters
Four possible localization for mitochondrial enzymes
The Golgi Complex
Located near cell nucleus, consists of flattened,
membrane-bounded sacs (cisternae) forming a stack

Each stack has:

cis-face is an entry face - adjacent to ER to accept
incoming vesicles
trans-face is an exit face – points towards plasma membrane, produces vesicles for
forward flow
The function of the Golgi is to transport and process
secreted and membrane proteins from ER to the cell surface

Cisternae segregated into

convex ("cis"), medial
(middle), and concave
("trans") compartments.
• Cis – removal of mannose,
phosphorylation
• Medial – removal of mannose,
addition of N-acetylglucosamine
ER
• Trans – Removal of galactose,
cys addition of sialic acid
medial • TGN – addition of sialic acid,
trans Sorting
TGN
Vesicular Transport
The Endocytic Pathway
Endosomes and Lysosomes
Lysosomes
•Lysosomes are active in recycling the cell's organic material and in the intracellular
digestion of macromolecules.
•Lysosomes contain various hydrolytic enzymes that are capable of digesting nucleic
acids, polysaccharides, fats and proteins.
•The inside of a lysosome is acidic.
•In humans, a variety of inherited conditions can affect lysosomes. These defects are
called storage diseases and include Pompe's disease and Tay-Sachs disease. People with
these disorders are missing one or more of the lysosomal hydrolytic enzymes.
The Cytoskeleton
 Cytoskeleton is a network of protein
filaments in the cytoplasm

Main functions:
• Supports large volume of the cytoplasm
• Participates in large-scale movements associated
with the changes in cell shape and cell motility
• Provides machinery for organelle transport,
chromasome segregation during mitosis, and cell
division
 Major components of cytoskeleton

Actin filaments Microtubules

Intermediate filaments
 The cytoskeletal filaments
• Common Features :
– Linear polymers of protein subunits
• Actin ( ~8 nm in diameter)
• Intermediate Filaments ( ~10 nm in diameter)
• Microtubules ( ~24 nm in diameter)
– Filaments are dynamic, i.e. they can assemble
and disassemble
– Highly conserved
Intermediate Filaments
• Intermediate filaments enable
cells to withstand mechanical
stress when cells are stretched.
• They can span the entire
cytoplasm and are anchored to
the plasma membrane.
The Microtubule Cytoskeleton

• Also penetrates the entire

volume of the cell

• Whereas actin fibers are

concentrated at the periphery,
most microtubules radiate from
a central location in the cell

• Main functions: intracellular

transport and mitosis
 Microtubules

Microtubules provide an
organizational structure in an
interphase cell and separate
chromosomes in a dividing cell

http://www.circs.neu.edu/external/Frank.Gibbons/spindle.html
Microtubules Provide Tracks for Transport
Microtubules are long hollow cylinders made of tubulin

•Protofilaments are linear chains of tubulin

dimers, a parallel bundle of 13 protofilaments
forms a microtubule
•There are three kinds of tubulins, each with
many subtypes:
α-tubulin and β-tubulin form α/β tubulin
dimers and represent the basic building block of
microtubules
γ-tubulin is involved in more specialized
processes, such as nucleation
Microtubules have a GTP “cap” stabilizing the
ends.
 Motor
Proteins
• Motor proteins bind to
microtubules and move
by cycles of
conformational changes
using energy from ATP.

• One end of the protein

can bind to specific
cellular components.
Actin filaments = microfilaments

•Actin is the most common protein in

the cytoplasm

•Actin filaments are concentrated

beneath the plasma membrane and
give the cell mechanical strength

•Assembly of actin filaments can

determine cell shape and cause cell
movement

•Association of actin filaments with

myosin can form contractile structures
Spectrin
Principal component of the cytoskelton
(protein meshwork underlying surface of
the red cell)

–Maintains structural integrity of the red

cell (e.g. biconvave shape)
–Long thin flexible rod
–Necessary as red cells go through small
capillaries