Cell

- basic structural and functional units

- smallest living parts of the body

Animal cells
- enclosed by cell membrane - eukaryotic
- with nucleus surrounded by cytoplasm - fluid containing a system of:
- membranous organelles
- non-membranous assemblies
- cytoskeleton
Bacteria
- prokaryotic
- have cell wall
- lack nuclei and membranous cytoplasmic structures

Average adult human body - nearly 40 trillion cells, derived from:

- zygote
- single cell: formed by merger of sperm with oocyte at fertilization

Blastomeres
- cells produced in the first zygotic cellular divisions
- part of the early embryo’s inner cell mass
- give rise to all tissue types of the fetus

Embryonic stem cells

- Explanted to tissue culture cells of the inner cell mass

Differentiation
- cells of the fetus undergo this specialization process
- express sets of genes that mediate specific cytoplasmic activities — becomes organized
in tissue and change shape accordingly.
e.g
Muscle cell precursors
- elongate into long, fiber-like cells containing actin and myosin
- are specialized for using these proteins to convert chemical energy into forceful
contractions.

Changes in cell’s microenvironment

- same cell type have variable feature and activities
Cells appearing similar structure
- have different families of receptors for signaling molecules
e.g
Breast fibroblasts and Uterine smooth muscle cells - sensitive to female sex
hormones
Fibroblasts and Smooth muscle cells - insensitive

Plasma Membrane
- cell membrane or plasmalemma
- envelopes eukaryotic cell consists of phospholipids, cholesterol, and proteins with
oligosaccharide chains
- selective barrier regulating the passing of material
- facilitates transport of molecules - site where materials are exchanged bet. cells and
environment.
- keep contant the ion content of cytoplasm
- defines outer limit of the cell
- 7.5. to 10 nm thick in diameter
- Visible only in electron microscope
- In a light microscope, seen faintly. Consists plasma membrane proteins + EC
material
- In Transmission Electron Microscope (TEM), trilaminar appearance after fixation
in osmium tetroxide
- Osmium binds polar head of phospholipids and oligo
chains
- produce two dark (electron dense) outlines that enclose a
light band of osmium-free FA (electron lucent band).

FUNCTIONS OF PLASMA MEMBRANE:

1. Physical barrier
- flexible boundary
- protect cellular contents
- supports cell structure

Phospholipid bilayer
- separates substances inside and outside the cell

2. Selective permeability
- regulates entry and exit of ions etc.

3. Electrochemical gradient
- establishes and maintains electrical charge difference

4. Communication
- contains receptors - recognize and respond to molecular signals
Membrane proteins
- perform specific recognition and signaling functions
- are less mobile
- involved in transduction of signals outside the cell
- such proteins are located in specialized membrane patches called
lipid rafts (high conc. of cholesterol and Saturated FA - reduce
lipid fluidity)
- with scaffold proteins, maintains spatial relationship bet.
enzymes and signaling proteins.
- also allows proteins to remain in close proximity and
interact more efficiently.

Integrins
- a plasma membrane protein
- linked to both cytoplasm and ECM
- allow continuous exchange of influences bet. cytoplasm and ECM

Membrane phospholipids
- are amphipathic
- Two non-polar long-chain fatty acids (hydrophobic or water repelling) linked to a charged
pole head (hydrophilic or water attracting) bears phosphate group
- most stable in double layer (bilayer)
- Hydrophobic FA: middle (away from water)
- Hydrophilic head: outer (contacting the water)

Tails of phospholipids (hydrophobic)

- Unsaturated FA: bent/kinked
- Saturated FA: straight

Sterol lipid (molecules of cholesterol)

- insert in the phospholipid
- restrict movements
- modulate fluidity of all membrane components

In red blood cells,

- Phosphatidylcholine and sphingomyelin - abundant in outer half
- Phosphatidylserine and phosphatidylethanolamine - inner layer
Glycolipids
- outer layer’s lipids
- include oligosaccharide chains that extend outward
- contribute to glycocalyx
- delicate cell surface coating
- fuzzy material on the outer surface of the membrane

Proteins
- major constituents of membrane (50% weight in plasma membrane)

Integral proteins
- within the lipid bilayer
- extracted: detergents - disrupt lipids

Peripheral proteins
- bound to one of the two membrane surfaces (cytoplasmic side)
- extracted: salt solutions

Multipass proteins
- polypeptide chains of integral proteins spanning the membrane

Transmembrane proteins
- completely span the bilayer

Freeze-fracture electron-microscope
- studies of membranes
- shows that integral proteins protrude from outer or inner membrane surface

Carbohydrate moieties of glycoproteins

- project from the external surface of cell membrane
- components of proteins as receptors
- cell adhesion, cell recognition, response to protein
hormones

All membranes in the cell are ASYMMETRIC

Fluid Mosaic model

- movable mosaic within the fluid lipid bilayer
- Proteins are not bound in place and move laterally
- restricted by cytoskeletal attachments
 Epithelial cells tight junctions
- restrict lateral diffusion of unattached transmembrane proteins and outer layer
lipids

Transmembrane Proteins and Membrane Transport

1. Diffusion
- transports small, nonpolar (uncharged) molecules directly inyo lipid bilayer
- Lipophilic molecules (fat-soluble) diffuse through membranes

2. Channels
- are multipass proteins
- forms transmembrane pores where ions pass selectively
- cells open and close specific channels for:
- Na
- K
- C
- and other ions; in response to physiological
stimuli

Aquaporins
- a channel protein where water molecules cross

3. Carriers
- are transmembrane proteins
- bind small molecules and translocate then across the membrane via
conformational changes
——————————————————————————————
Diffusion, channels, and carriers
- allows movement of substances across membranes down a concentration gradient due
to its kinetic energy

Membrane pumps
- are enzymes engaged in active transport
- utilize energy from hydrolysis of ATP to move ions and other solutes across membranes
- consume ATP pumps so they are called ATPases

Transport by Vesicles: Endocytosis and Exocytosis

Endocytosis
- Macromolecules enter cells by being enclosed within the folds of plasma membranes
which fuse and pinch off internally as spherical cytoplasmic vesicles or vacuoles.
THREE MAJOR TYPES OF ENDOCYTOSIS:
1. Phagocytosis
- “cell eating”
- ingestion of particles
- macrophages and neutrophils are specialized for this activity

Phagosome
- fusion of membranous folds encloses the bacterium in this intracellular vacuole
- merges with a lysosome for degradation

2. Pinocytosis
- “cell drinking”
- smaller invaginations of cell membrane which duse and entrap EC fluid

Pinocytotic vesicles
- 80 nm in diameter
- pinch off inwardly from the cell surface and either:
- fuse with lysosomes
- move to the opposite cell surface where they fuse with membrane

Transcytosis
- bulk transfer of dissolved substances across the cell

3. Receptor- mediated endocytosis

- includes membrane proteins called receptor
- bind specific molecules (ligands)
Integral membrane proteins
- receptors of many substances (LDL and protein hormones) at the cell
surface
- High affinity causes these proteins to aggregate in special membrane
regions and invaginate and pinch off internally as vesicles

Peripheral proteins
- formation and fate of vesicles in receptor-mediated endocytosis
- Occupied cell surface receptors associates and begin to invaginate as
coated pits

Clathrin
- polypeptide on the electron dense coating on the cytoplasmic
surface of coated pits.
- forms a region of cell membrane into cage-like invagination that
soon pinches off into the cytoplasm as a coated vesicle.
 Caveolae
- L. caveolae or little caves
- invaginations prominent in very thin cells
- involved in family of integral membrane proteins: caveolins
- associated with diverse peripheral proteins: cavins

Endosomal compartment
- dynamic collection in the peripheral cytoplasm of membranous tubules and vacuoles

Rab proteins
- directs vesicle trafficking through endosomal compartment
- peripheral membrane G-proteins
- small GTPases that bind guanine nucleotides and associated proteins.

Exocytosis
- movement of of large molecules rom inside to outside the cell
- molecules to be secreted fuses with plasma membrane → release of its contents into the
extracellular space
- triggered by a transient increase in cytosolic Ca2+.

TWO PATHWAYS OF EXOCYTOSIS:

1. Constitutive Secretion
- used for products that are release from cells continuously
e.g. collagen subunits for ECM

2. Regulated Secretion
- occurs in response to signal coming to the cells
e.g. release of digestive enzymes from pancreatic cells in response to a specific
stimulus.
- From epithelial cells, occurs at the apical domains of cells → glandular
secretion

Endocytosis → cell membrane become part of endocytotic vesicles/ vacuoles

Exocytosis → returned to the cell surface

Membrane Trafficking
- membrane movement and recycling
- occurs continuously in most cells
- crucial for maintaining cell
- reduce blood lipid levels

Multivesicular bodies
- accumulate small vesicles within their lumens
 - may merge with lysosome for degradation
- may fuse with plasma membrane → release the intraluminal vesicles outside the cell
called exosomes (<120 nm in diameter)
- can fuse with other cells transferring their contents and membranes

Signal Reception and Transduction

Gap Junctions
- couple cells
- allow exchange of ions and small molecules

Target cells
- cells bearing receptors for a specific ligand

Signaling process:
1. Endocrine signaling
- Hormones are carried in the blood throughout the body

2. Paracrine signaling
- chemical ligand diffuses in EC fluid but is rapidly metabolized

3. Synaptic signaling
- special kind of paracrine signaling
- Neurotransmitters act on adjacent cells through special contact areas called
synapses.

4. Autocrine signaling
- signals bind receptors on the same cells that produced the messenger
molecule

5. Juxtacrine signaling
- Cell membrane-bound proteins bind surface receptors of the target cell when
two cells make direct physical contact

Receptors for hydrophilic signaling molecules

- Polypeptide hormones and neurotransmitters
- are usually transmembrane proteins in the plasmalemma of target cells
THREE IMPORTANT FUNCTION CLASSES OF RECEPTORS:
1. Channel-linked receptors
- promote transfer of molecule or ions across the membrane
- bind ligands such as neurotransmitters and open to allow influx of specific ions

2. Enzymatic receptors
- induces catalytic activity in associated peripheral proteins
- usually protein kinases that activated to phosphorylate other proteins

3. G-protein - coupled receptors

- stimulate associated G-proteins → bind guanine nucleotide GTP
→ released to activate cytoplasmic proteins
- change the conformation of G protein subunit → binds to GTP → activates and
release protein such as ion channels and adenylyl cyclase

Pseudohypoparathyroidism → caused by non functioning parathyroid

Dwarfism → growth hormone receptors
- In these two conditions, the gland produce the respective hormones but target cells
cannot respond because they lack normal receptors

Signal transduction
- Ligands binding such receptors in a cell membrane → first messengers
- activates series of intermediary enzymes to produce changes in cytoplasm

Channel-mediated ion influx/ activation of kinases

- activate various cytoplasmic proteins, amplifying the signal

Activated G-proteins
- target ion channel or other membrane bound effectors that propagate the signal further
into the cell
- Adenyl cyclase → generates large quantities of second messenger molecules such as:
- Cyclic adenosine monophosphate (cAMP)
- 1,2 -diacylglycerol (DAG)
- inositol 1, 4, 5 - triphosphate (IP)
- Ionic changes or second messengers
- amplify first signal
- trigger enzymatic activity (usually kinases) → changes in
gene expression or cell behavior
- may diffuse in cytoplasm
- retained locally by scaffold proteins
Cytoplasmic Organelles
Fluid cytoplasm/ cytosol
- inside the cell membrane, it bathes organelles.
- metabolically active structures
- may be membranous (e.g. mitochondria)
- or non-membranous (e.g. ribosomes and proteasomes)
- position in the cytoplasm by movements along the polymers of
cytoskeleton.
- determines a cell’s shape and motility
- contains hundreds of enzyme (such as of the glycolytic pathway), which:
- produce building blocks for larger molecules
- break down small molecules to liberate energy.

Ribosomes
- macromolecular machines
- 20 x 30 nm in size
- assemble polypeptides from amino acids on tRNA in a sequence specified by mRNA
- has two subunits different in size:
1. small ribosomal subunit
- highly folded rRNA chain
- associated with more than 30 unique proteins

2. large subunit
- has three other rNA molecule
- nearly 50 other basic proteins

- rRNA molecules in ribosomal unit

- provide structural support
- position tRNA molecules bearing amino acids in the correct “reading frame”
- catalyze the formation of peptide bonds

- Peripheral proteins of ribosome

- stabilize the catalytic RNA core
- synthesized in cytoplasmic ribosomes → imported to the nucleus (where they
associate with newly synthesized rRNA) → (ribosomal units formed) move from
the nucleus → cytoplasm (ribosomale units are reused for translation of any
mRNA strand)

- During protein synthesis, ribosomes bind to the same strand of mRNA to form larger
complexes called polyribosomes or polysomes
- In stained preparations of cells, are intensely basophilic because of numerous
phosphate group of the constituent RNA molecules that act as polyanions.
 - Cytoplasmic regions that stain intensely with hematoxylin and basic dyes
(methylene and toluidine blue) indicate sites of active protein synthesis.
- Proteins are made in the polysomes attached to ER that are to be:
- incorporated into membranes
- stored in lysosomes
- secreted from cell

Protein chaperones
- guides proper folding of new proteins

Denatured proteins
- those that cannot be refolded properly
- conjugated to the protein ubiquitin
- targets them from breakdown by proteasomes (proteasomal degradation)

Isolated cytoplasmic clusters

- are polyribosomes where proteins are synthesized:
- for use within the cytosol
- for import into the nucleus and certain other organelles

Polyribosomes attached to membranes of ER

- translate mRNA coding for membrane proteins of ER, Golgi, or cell membrane
- enzymes to be stored in lysosome
- proteins to undergo exocytosis from secretory vesicles

Free polyribosomes
- synthesize:
- cytosolic and cytoskeletal proteins
- proteins for import into the nucleus, mitochondria, and peroxisomes

Endoplasmic Reticulum
- convoluted membranous network (anastomosing network)
- extends from the surface of the nucleus throughout most of the cytoplasm
- encloses intercommunicating channels called cisternae (L. cisternae, reservoirs)
- formed by continuous membrane
- membrane surface up to 30 times the plasma membrane
- major site for vital cellular activities
- biosynthesis of proteins and lipids
- for synthesis, transport (moves molecules through cisternal space), storage (for newly
synthesized molecules), and detoxification
TWO TYPES OF ER:
Rough Endoplasmic Reticulum
- prominent in cell specialized for protein secretion:
- pancreatic acinar cells (making digestive enzyme)
- fibroblasts (collagen)
- plasma cells (Ig)
- site for synthesis of most membrane-bound proteins
- synthesizes proteins for secretion, incorporation into the plasma, membrane, and as
enzymes within lysosomes
- MAJOR FUNCTION:
- production of membrane associated proteins
- proteins of membranous organelles
- proteins to be secreted by exocytosis
- mediated by resident enzymes of RER and
protein complex
- acts as chaperones guiding the folding of nascent
proteins
- inhibiting aggregation
- monitor protein quality within ER
- cisternae are flattened
- limited by membranes continuous with outer membrane of nuclear envelope
- Presence of polyribosomes: confers basophilic staining properties when viewed in light
microscope

Smooth Endoplasmic Reticulum

- for lipid biosynthesis, detoxification of harmful compounds, sequestration of Ca2+ ions.
- site for lipid synthesis and carbohydrate metabolism
- detoxify drugs and alcohol
- cisternae are tubular/ saclike with interconnected channels
- continuous with RER
- lack polyribosomes
- not basophilic
- best seen in TEM

THREE MAIN FUNCTIONS OR SER:

1. Enzyme of SER synthesize phospholipids and steroids → transferred to SER by:
- by lateral diffusion
- by phospholipid transfer
proteins
- by vesicles
- In cells that secrete steroid hormones, SER occupies large portion of cytoplasm

2. Cytochrome P450 family, allow detoxification of potentially harmful exogenous molecules

- In liver cells, these enzymes process endogenous molecules (components of bile)
 3. SER vesicles
- responsible for sequestration and controlled release of Ca2+
- functions in striated muscle cells
- SER has an important role in the contraction process and assumes a specialized form
called sarcoplasmic reticulum.

MEDICAL APPLICATION
Jaundice
- yellow discoloration of the skin
- accumulation in EC fluid of bilirubin and other pigments which are normally metabolized
by SER

Protein synthesis
- begins on polyribosomes in cytosol
- Protein synthesis of RER location:
- intracellular storage (lysosomes and leukocytes)
- provisional storage in cytoplasmic vesicles (pancreas and endocrine cells)
- as integral membrane proteins

Signal Recognition Particle (SRP)

- a protein complex where newly translated signal sequence are bounded
- inhibits further polypeptide elongation
SRP-ribosome-nascent peptide complex
- binds to SRP on ER
- SRP release signal sequence → translation with nascent popypeptide chain
transferred to translocator complex (also called translocon)
- Inside the lumen of RER, signal sequence is removed by signal peptidase.
- With ribosomes on ER surface, translation continue
- Chaperone and other proteins pull nascent polypeptide through
the translocator complex.
- Upon release from ribosome, post translational modification and proper folding of
the polypeptide continue

ER-associated degradation (ERAD)

- new proteins that cannot be folded or assembled by chaperones undergo this process.
- unsalvageable proteins are:
- translocated back to cytosol
- conjugated to ubiquitin
- and then degraded by proteasomes

Golgi Apparatus
 - Golgi complex
- dynamic organelle
- completes post translational modifications of proteins produced in RER
- packages and addresses proteins produced in RER to their proper destinations
- Named after Camillo Golgi in 1898
- consists of smooth membranous:
- saccules
- vesicular
- flattened
- all contains enzymes and proteins being processed
- located near the nucleus
- has two face:
1. cis face
- receiving region
2. trans face
- shipping region
- where larger saccules or vacuoles accumulate, conense, and
generate other vesicles that carry completed protein products to
organelles away from Golgi.

Transport vesicles
- a small membrane-enclosed carriers where material moves from the RER cisternae to
Golgi
- are transported along cytoskeletal polymers by motor proteins
- merge with cis face

Formation of transport vesicles and secretory vesicles

- driven by coat proteins (including clathrin)
- regulate vesicular traffic through and beyond Golgi

Coat protein COP - II

- promotes forward movement of vesicles in cis golgi network of saccules

COP - I
- retrograde movement in that region

The Golgi apparatus

 - high plastic, morphologically complex system of membrane vesicles and cisternae in
which proteins and other molecules made in the RER undergo further modification and
sorting into specific vesicles destined for different roles in the cell.
- initiates packaging, concentration, and storage of secretory products
- In TEM, provide evidence how this organelle works
- In SEM, morphological aspects of Golgi are revealed clearly.
- It show a three-dimensional snapshot of the region between RER and the Golgi
membrane compartments
- In trans Golgi network, proteins and glycoproteins combine with specific receptors
- Location: seen clearly in intact cultured cells processed by immunocytochemistry using
antibody against golgin-97
- Enzymes: important for glycosylation, sulfation, phosphorylation, and limited proteolysis
of proteins

Transport vesicles
- move proteins serially through the cisternae until at the trans face or shipping region

Inset
- a small region of a Golgi apparatus in a 1 um section from a silver-stained cell
- demonstrates glycoproteins within cisternae

Secretory Granules
- condensing vesicles in the Golgi apparatus
- found in cells that stora product until its release by exocytosis
- signaled by a metabolic, hormonal, or narula message (regulated secretion)
- 200 times more concentrated than the cisternae of RER

Zymogen granules
- secretory granules with dense contents of digestive enzyme

Lysosomes
- Greek, lysis (solution) + soma (body)
- spherical
- 0.05 to 0.5 um in diameter
- sites of intracellular digestion
- turnover of cellular components
- optimal activity at acidic pH (5.0)
- leaked lysosomal enzyme are inactive at pH of cytosol (7.2)
- membrane limited vesicles with 40 different hydrolytic enzymes
- abundant in cells with great phagocytic activity (macrophages, neutrophils)
- In light microscope, slightly larger and visible after histochemical staining
- In TEM, uniform granular electron dense appearance
 - Nature and activity depends on cell type
- Most common: acid hydrolase
- proteases, nucleases, phosphatases,
phospholipases, sulfatases, B-glucuronidase
- digest organelles/ membrane by autophagy

Lysosomal hydrolases
- synthesized and segregated in the RER
- transferred to Golgi
- enzymes are further modified and packed in vacuoles → lysosome

Marker mannose-6-phosphate (M6P)

- added by phosphotransferase in cis Golgi to N-linked oligosaccharides of hydrolases
- In trans Golgi, bind M6P-containing proteins and divert them from the secretory pathway
for segregation into lysosomes

- Material taken from outside the cell by endocytosis is digested phagosome/pinocytotic

vesicle fuses with a lysosome
- mixes endocytosed material with lysosomal enzyme
- activates proton pumps in the lysosomal membrane → acidify contents, allowing
digestion
- composite, active organelle is now termed as secondary or hetero lysosome
- larger and have more heterogeneous appearance in TEM

Residual Body
- a small vascular remnant where indigestible material is retained
- In long lived cells (neurons, heart muscle), it can accumulate as granules of lipofuscin

Autophagy
- excess organelles or large aggregates of nonfunctional macromolecules in cytoplasm
are degraded
- lipid bilayer forms around and isolates the cytoplasmic portion to be removed producing
autophagosomes
- Greek, autos (self) + phagein (to eat) + soma
- fuses with lysosome for digestion of enclosed material
- formed after nonfunction or surplus organelles become enclosed
with membrane
- resulting structure fuse with lysosome
- enhanced during starvation or nutrient stress
- Released from autophagosomes for reuse as sources of energy or in new synthetic
reactions:
- amino acids
- nucleotides
- other digestion products
Phagocytosis of bacteria or debris: by neutrophils
Secretion of hydrolytic enzymes: by osteoclasts

Endocytosis produces vesicles

- fuse with endosomes → merge with lysosomes

Phagocytic vacuoles (phagosomes)

- fuse with primary lysosome = secondary lysosomes or hetero lysosomes
- where ingested material is degraded

Proteasomes
- very small abundant protein complexes
- size of a small ribosomal unit
- degrade denatured or nonfunctional polypeptides
- remove proteins no longer needed
- restrict activity of specific protein in a certain time
- deal primarily with free proteins as individual molecules

- cylindrical
- made of four stacked rings, each composed with seven proteins (including proteases)
- at the end of cylinder: regulatory particle containing ATPase
- recognizes proteins with attached molecules of ubiquitin
- abundant cytosolic 76-amino acid protein found in
cells

- Misfolded proteins are recognized by other complexes that

conjugate ubiquitin to lysine followed by formation of
polyubiquitin chains.

- Ubiquitinated proteins
- recognized by regulatory particles of
proteasomes
- unfolded by ATPase using energy from ATP
- translocated to the core of cylindrical
structure
- degraded by endopeptidases

- Ubiquitin molecules
- released for reuse
- peptides produced may be broken to amino
acids
MEDICAL APPLICATION:
Failure of proteasomes
- protein accumulates → damage/ kill cells → lead to neurodegeneration → Alzheimer’s
disease and Huntington’s disease

Mitochondria
- Greek, mitos (thread) + chondros (granule)
- elongated structure
- 0.5 -1 um diameter
- length up to 10 times greater
- high plastic (same with Golgi)
- rapidly changing shape
- moved through cytoplasm along microtubules
- where protein synthesis occurs
- membrane enclosed organelles with arrays of enzymes specialized for:
- AEROBIC respiration
- production of ATP - supplies energy for cellular activities
- Enzymes: yield 15 times more ATP tna produced by glycolysis alone
- some energy released in mitochondria is not stored in aTP but as heat to
maintain body temperature
- Number of mitochondria is related to cell’s energy needs;
- high energy metabolism = abundant mitochondria
- In TEM, have two separated and very different membrane that create two compartments:
1. Innermost matric
2. Intermembrane space
- both contains higher density of protein molecules
- have reduced fluidity
- Outer membrane
- sieve-like
- smooth
- contain transmembrane proteins called porins
- forms channel through which small molecules such as
pyruvate pass from the cytoplasm to the intermembrane
space
- Inner membrane
- has many sharp folds or long folds called cristae
- project into the matric
- greatly increase memrabrane’s surface area
- number depends to the energy need of cells

- Lipid layer of Inner membrane

- contains unusual phospholipids
- highly impermeable to ions
 - Matrix
- gel with high concentration of enzymes

- Integral proteins
- transport proteins that make the inner membrane selectively permeable
to small molecules
- Mitochondrial matrix enzyme
- oxidize pyruvate and FA to form acetyl coA

Glycolysis
- converts glucose ANAEROBICALLY to pyruvate in cytoplasm
- pyruvate imported to mitochondria → oxidized to CO2, and H2O

- Enzymes embedded in inner membrane allow oxidative phosphorylation

→ produce ATP in animal cells

Chemiosmotic process
- Formation of ATP by oxidative phosphorylation enzymes
- Protons accumulating in the intermembrane space = Electrochemical gradient

Membrane- associated proteins of ATP synthase system

- forms large (10 nm), multisubunit, globular complexes on stalk-like structure
- project from the matrix side of the inner membrane

Apoptosis
- In the cytoplasm, cytochrome c activates sets of proteases that degrade all cellular
components in this process.
- results in rapid cell growth

Peroxisomes
- spherical organelles
- enclosed by single membrane
- produce and degrade hydrogen peroxide H2O2
- formation of bile acids and cholesterol

Oxidases
- oxidize substrates by removing hydrogen atoms that are transferred to molecular
Oxygen = H2O2
Catalase
- breaks down H2O2 to water and O2
- inactivate potentially toxic molecules (in liver and kidney cells)

B-oxidation of long chain FA

- 18 carbons and longer
- accomplished y peroxisomal enzyme

PEROXISOMES FORMS IN TWO WAYS:

1. Budding of precursor vesicles from the ER
2. Growth and division of preexisting peroxisomes

Peroxisomal Proteins
- synthesized on free polyribosomes
- have targeting sequence of amino acids

MEDICAL APPLICATION:
Neonatal adrenoleukodystrophy
- defective integral membrane protein needed for transport of long FA into peroxisome for
- oxidation
- Accumulation can disrupt myelin sheath in nerve tissue

Zellweger syndrome
- affects structure and function of several organ system

Cytoskeleton
- complex array of:
- microtubules
- fine tubular structure
- highly dynamic in length
- organized into larger, more stable arrays called axonemes in the
cytoplasmic extensions called cilia and flagella
- have DYNAMIC INSTABILITY
- microtubules of cilia: stabe
- mitotic spindle: short-lived
- 25 nm in diameter; wall is 5 nm thick
- Location: cytoplasm at centrosomes; axonemes
- Functions:
- Maintain cell shape and polarity
- provide tracks for organelle and chromosome movement
- move cilia and flagella
- Heterodimer of a and B tubulin
- protein subunit of microtubule
- molecular mass of 50 kDA
 - polymerize to form microtubules (have slight spiral organization)
- align lengthwise as protofilaments
- Microtubule Organizing Centers (MTOCs)
- short assemblies of tubulin
- act as nucleating sites for further polymerization
- energy for assembly is derived from GTP
- Dominant: centrosome
- organized around two cylindrical centrioles
- 0.2 um in diameter; 0.3 - 0.5 um in length
- composed of nine highly organized
microtubule triplets
- have long axes at right angles

- In DNA replication, centrosome duplicates itself

→ each centrosome has two pairs of centrioles

- In mitosis, divides into have → to opposite pole

Motor proteins
- Transport along microtubules is under the control of this protein
- use ATP in moving

Kinesins
- carry material away from the MTOC reach the nucleus
- toward the plus end of microtubules (anterograde transport)

Cytoplasmic dyneins
- along microtubules in the opposite direction (retrograde transport)
- toward the nucleus

- microfilaments (actin filaments)

- 5 - 7 nm in diameter
- more flexible than microtubules
- composed of G-actin monomers
- Arp ⅔ binds to the site of preexisting actin filament and induces a new
F-actin branch
- a process which can lead to formation of microfilament network
- HIGHLY DYNAMIC
- Location: beneath cell membrane; in cell extensions like microvilli

- Functions:
- allow motility and most contractile activity in cell
- contract and move cells
- change cell shape
 - cytoplasmic transport and streaming

Thread Milling
- a process of migration of subunit though polymer
- assembly and disassembly of subunits from F-actin are promoted by profilin and cofilin

Lamellipodia
- cell movements on firm substrate involve in this sheet-like protrusions

Cell cortex
- cytoplasmic region

Cross linking within network of F-actin

- increases cytoplasmic viscosity

Severing and capping filaments

- decrease viscosity

Myosin motors
- use ATP to transport cargo along F-actin
- Movement: towards the plus end of actin filaments
- Myosin VI is the only known myosin that moves in the other direction

Interaction between F-actin and myosins form the basis for various cell movements:
- Transport of organelles, vesicles, and granules in the process of cytoplasmic streaming
- myosin II constricting to produce two cells by cytokinesis during mitosis
- myosin I moves along microfilaments to produce the cell surface changes during
endocytosis

- intermediate filaments
- 8-10 nm in diameter
- STABLE
- Location: throughout cytoplasm; at desmosomes; inside nuclear envelope
- Functions:
- strengthen cell and tissue structure
- maintain cell shape
- maintain nuclear shape (lamins)
- Proteins ranging size in 40 to 230 kDA serve as subunits of various
intermediate filaments

Antiparallel tetramers
- coiled, rold-like dimers
- self-assemble into large cable-like bundles or protofibrils
 - stabilized by further lateral interaction

- determine the cell shape

- role in movements of organelles and cytoplasmic vesicle
- allow movement of entire cells

Intermediate filament proteins with particular biological, histological, or pathological importance:

1. Keratins
- Greek, keras (horn)
- diverse family of acidic and basic isoforms
- compose heterodimer subunits of intermediate filaments in all epithelial cells
- produce filaments
- form large tonofibrils that attach to certain junctions between epithelial cells
- cytokeratins accumulate during differentiation in the process of keratinization
- produce outer layer of nonliving cells
that reduced dehydration
- provides some protection from minor
abrasions
- produces various hard protective
structures of skin

2. Vimentin
- most common class III intermediate filament protein
- found in cells derived from embryonic mesenchyme
- Desmin found in muscle cells
- Glial fibrillary acidic protein (GFAP) found in astrocytes
- supports cells of CNS tissue

3. Neurofilament
- make heterodimers
- form the subunits of the major intermediate filaments of neuron

4. Lamins
- family of seven isoforms present in the cell nucleus
- form structural framework called nuclear lamina

Inclusions
- containing accumulated metabolites
 - have little or no metabolic activity
- transitory structures NOT enclosed by membrane

Features important and commonly seen:

1. Lipid droplets
- accumulations of lipid-filling adipocytes (fat cells)

2. Glycogen granules
- aggregates of carbohydrate polymer in which glucose is stored
- visible as irregular clumps of periodic-acid Schiff (PAS)
- In liver cells, positive or electron dense material

3. Melanin
- dark brown granules
- In skin, serve to protect cells from ultraviolet radiation

Lipofuscin
- pale brown granule stable in nondividing cells
- contains complex mix of material partly derived from residual bodies after
lysosomal digestion

Hemosiderin
- dense brown aggregate of denatured ferritin proteins with many atoms bound
iron
- prominent in phagocytic cells of liver and spleen

MEDICAL APPLICATION:
Hemosiderosis
- iron containing inclusion hemosiderin occurs in cells of organs
- increased uptake of dietary iron

Hemochromatosis and Iron overload syndrome

- extreme accumulations of iron in cellular hemosiderin
- liver and organs are damaged