1.
DIRECT ATP-DEPENDENT Active transport
-depend on ATP particularly the hydrolysis of the ATP
-breakage in the phosphoanhydride bond releasing ADP
and Pi
If you take a look at the carrier protein or the PUMP
involve in active transport
PUMP has two functions:
a Transport-carry out the translocation of their
substrates or solute against its concentration
gradient
a ATP hydrolysis -enzyme to catalyzes ATP hydrolysis
So the cation pumps that we know of-
Sodium potassium pumps
ATPase pumps
Ca2+ ATPase pumps
Classification of these pumps:
1 P class
-get phosphorylated
-means after binding ATP, hydrolyzing it, they
release ATP back into cytosol, but Pi may bound to
the pump
-phosphorylation triggering change in conformation
-NOTING THAT CARRIER PROTEIN works by changin g
conformation
e.g. Na+K+ ATPase, Ca2+ ATPases
*ATPase-enzymes that catalyzes ATP and also transport
1 V class
-Not phosphorylated
When they hydrolyze ATP, they released both ATP
and Pi back into the cytosol
a V1 domain-peripheral membrane proteins carrying
out ATP hydrolysis
a V0 domain-integralmembrane proteins responsible
carrying out active transport usually of H+
e.g. proton pumps such as H pumps
*Found in the membrane in vacuoles in plant and
fungal membranes, and membranes in lysosomes and
endosomes of animal cells.
*Lumen are acidic, hence, high [H+ ions ]
*PUMPS-actively transport H+ ions from the cytosol into
the lumen
-regulate pH in the cytosol so it wouldn't be acidic
-responsible for procuring excess H+ ions bringing
them back inside the lumen of the acidic organelle
1 ABC superfamily (ATP Binding Cassettes)
a Two T domains- transmembrane/integral membrane
proteins
-responsible for actual translocation of
substrates/solute
a Two A domains- peripheral membrane proteins
-responsible binding and hydrolyzing ATP
SUPERFAMILY because made up of several hundred
membranes/types of carrier proteins
*Find in wide range of organisms:
bacteria, plants, fungi and humans
*Substrates they carry depending on the carrier
proteins:
Ions, polysaccharides, amino acids, lipids, peptides
even proteins,
*Directionality of transport: some inside going out,
outside going in
VARIATION: what they transport and direction which
they transport
SPECIFIC EXAMPLES:
Na+K+ ATPase pump
Classification:
Co-transport
Antiport-opposite
P-class antiport
Electrogenic-function is to establish
voltage/membrane potential
Review: in terms of concentration gradient,
more Na+ outside the cell, K+ ions inside
Difference in concentration 100 mM
-responsible for maintaining the concentration gradient
of Na+
SIGNIFICANCE:
a store E needed for other metabolic processes in
the cell/ other type of active transport (Sodium
Motive Force)
a Do prevent cell swelling in isotonic medium
i Osmolarity in the cell is ensured to not
increase therefore preventing the entry of cell,
hence preventing cell swelling
2 CONFORMATIONS: cycle can start anywhere
a E1-high affinity for 3 Na+ ions
 -open on the cytosolic side
a E2- low affinity for 3 Na+ ions but high affinity for
K+ ions
Ex. The pump is at an
a E1 state- at this point, it has high affinity for 3
Na+ ions and 1 ATP molecule so it bind
b ATP hydrolysis-
P class so phosphate is remain bind to the
pump only ADP is released to the cytosol
a E1 becomes E2- due to change in conformation so
-3 Na+ released to the extracellular fluid, and 2.INDIRECT/SECONDARY ATP DEPENTDENT ACTIVE
high affinity for 2 K+ ions so it will bind, after
TRANSPORT
the binding they are de-phosphorylated
releasing the phosphate causing change in
conformation going back to
a E1 state- low affinity for the K+ ions so they will be
released in the cytosol and pick up 3 Na+ from the
cytosol also
-3 Na+ outside, K+ inside
*Delivered to where they are more in
concentration
Ca2+ ATPase Pump
Remember the SR, sarcoplasmic reticulum found in the
2+
muscle cell containing a lot of Ca ions, has Ca 2+
release channels in the membrane that when opened
2+
because of opening of Ca voltage-gated channels in
the T tubule since they are physically connectedt and
due to change in voltage, they will open up causing
release of a lot of Ca2+ ions in the muscle cell cytosol
initiating muscle contraction
Need to relax, Ca2+ ions should be relax to the SR
Problem: SR still has moreCa2+ inside than cytosol
2 States:
2+
a E1-high affinity for 2 Ca ions
-cytosolic gate is open
-lumenal portion is closed
a E2-lost affinity for 2 Ca2+ions releasing them inside
the lumen
-cytosolic gate is closed
-lumenal portion is open
SR lumen has more Ca, cytosol has less Ca
In E1 state, Ca2+ ATPase Pump will pick up 2 Ca2+ ions
and then ATP binds and hydrolyzes and pump becomes
phosphorylated (P class) changing conformation E1 to
E2 because of that change, the 2 Ca2+ ions because
there is decrease in the binding affinity with the pump
will now be release into the lumen and eventually get
dephosphorylated so goes back to E1 to pick up another
2 Ca2+ ions from the cytosol. This goes on until the
concentration of Ca2+ ions in the cytosol normalizes , it
is reduce and muscle cell will now relax.
-does not directly depend on ATP hydrolysis but instead
in proton motive force or E stored in the concentration
gradient certain molecules (Na)
-carriers or pumps do not hydrolyze ATP because they
would come from proton motive force
e.g.active transport of glucose
Absorb glucose from small intestine (intestinal gut)
Na+- Glucose symport-transport Na + and glucose in
one directions)
More Na+ outside the cell, more glucose inside small
intestine cell compared to the gut
Carry out two types of transport
For every two Na+ ions , that are passively
transported that will (provide E) drive the
active transport of 1 molecule of glucose
inside the cytosol
Is Na+ meant to stay inside the cytosol of the small
intestine cell?
They should be circulated- move out of the cell-
diffuse across the blood vessels into the blood
stream and be circulated.
High concentration of+ glucose inside and
lower outside, when glucose move out of small
intestine cell via passive transport involving
glucose channel that passively transport
glucose from inside of the cell going to the
tissue fluid and diffuse into the blood, location
on the basal side of the cell
Na+ ions that were passively transported
during glucose uptake, move out of the cell in
 order to maintain the concentration gradient
of Na+. Keeping in mind that more Na is
outside so , r sodium potassium ATPase is
responsible in returning the sodium ions
outside the cell is active type of transport,.
Therefore, maintaining that concentration
gradient of sodium potassium so there will be
stored E/sodium motive force to carry out the
uptake of more glucose or active transport
of more glucose.
This is also known as ATP has an indirect role.
ATP somehow has a role , although indirect
role..
How so?
Immediate source of E here (glucose
uptake) is sodium motive force-E stored
in concentration gradient of Na.
How is that maintained?
Sodium Potassium ATPase pump. It
depends on ATP hydrolysis.
Without ATP, it cannot function, can't
maintain CG .
3.LIGHT-DRIVEN Active transport
-depend on light
-found in some archaeans- Halobacterium halobium
In their membrane bacteriorhodopsin is observed
-multipass integral membrane protein containing
chromophore (light absorbing organic group hence
sensitive to light
What happens when light activate bacteriorhopsin
-cause change in conformation from trans to cis
-accomplish the active transport of H+ ions
from inside going out
-PURPOSE: to maintain concentration gradient
of H (because it also stores E/proton motive
force)
4.GROUP TRANSLOCATION
-gram negative bacteria
-compound that is dephosphorylated is not ATP but PEP
-what happens is ,that solute or molecule transported is
chemically modified/phosphorylated
-Glucose ---> G-6-P
-BACTERIALPHOSPHOTRANSFERASES SYSTEM
e.g In cytosol, E1-catalyzes the hydrolysis of PEP, PEP
after hydrolysis releases pyruvate but E1 is
phosphorylated though (transient). Eventually, this
phosphate group is transferred to H protein, then H
protein will transfer it (phosphate group) to E2a (now
you have an idea why it is called the
phosphotransferases system) then phosphate group will
be transferred to E2b. Then you have this integral
membrane protein E2c that carries out the actual
transport of glucose from outside the cell going inside.
At the same time that it will transfer glucose from
outside going to the cytosol, it will also catalyze the
transfer of the phosphate group from E2b to glucose so
that in the end what is released into the cytosol is now
G6P.
With this, the glucose that is being chemically modify,
can it now get back out of the cell? Can it move out of
the cell using the same transport protein?
NO because this is just specific for glucose, there is
no transport protein that will recognize
G6P.Therefore, this system ENSURE THAT THERE IS
UNIDIRECTIONAL GRADIENT FOR GLUCOSE. It is
outside the cell, those phosphorylated will remain
inside the cell.
In bacteria, glucose is not the only sugar that can be
metabolized but also mannitol, mannose and lactose. So
they are also translocated via PTS. They also would
make use of E1 responsible for the first step-
hydrolyzezPEP. Also involve, H protein. However, E2a,
E2b and E2c will now be specific and different in
different sugars-they have similar functions but different
components.
MEMBRANE FLOW SYSTEMS
-TRANSPORT OF LARGE MOLECULE INVOLVING VESICLES
2 Main Stages of for endocytosis and exocytosis
1. Membrane adherence
-referring to different segments of the membrane
that will come close together or that will adhere to
each other.
-e.g In exocytosis, a vesicle with its own lipid
bilayer or membrane, will go near PM
In endocytosis, initially separate would come
in apposition with each other
 1 Membrane joining/fusion
-e.g.In exo, the two initially separate will now
fuse/join.
-gain more membrane material
In endo, fusion
-loss of segment in the PM
Rate should be equal of the two processes.
1.EXOCYTOSIS-process of secretion
-formed in Golgi apparatus (interior part of the cell)
In order to accomplish membrane adherence, it will
move at certain distance and go to the cell surface,
that movement is accomplished because the
secretory vesicles are carried by motor proteins
that are moving along microtubules. Motor proteins, 2.ENDOCYTOSIS -when cell internalize molecules (particularly
microtubules, are important for exocytosis because macromolecule from ECM)
they are responsible for making the secretory
vesicle move/travel a certain distance from the
Golgi apparatus area to the PM area.
a Secretory vesicle moves.
a Membrane adherence
a Rupture of part of the membrane
b Release of the contents of the secretory vesicles
a Fusion and integration of the vesicle lipid bilayer
into the PM
WHAT HAPPENS TO THE MATERIALS SECRETED?
They could either remain on the surface of the cell, like
membrane proteins to be inserted in to the
membrane/components of the cell wall
Or could be part of the ECM
If it is hormone, they should diffuse into the blood so it
will move and go to its specific target.
Types of exocytosis
1 Constitutive
e.g.constitutive gene expression
-synonym: CONTINUOUS/always
-the moment it is formed, it will now fuse with the
membrane and released its contents
1 Regulated
-formation of a secretory vesicle but will just
stay inside the cytosol, waiting for the
appropriate signal/stimulus before it will
release its contents into the cytosol
So if you recall our discussion in the motor
neurons and muscle contraction, there are
vesicles in the motor neurons containing the
neurotransmitter acetylcholine, remember that
those vesicles will only release the
acetylcholine into the synapse only when there
is an action potential that had reached the
terminus of the motor neuron axon initiating
the influx of Ca2+ ions and that is the only
time, that those vesicles would release/fuse to
the membrane and release acetylcholine into
the synapse.
-vesicles that are formed can move inward toward
the interior of the cell because of the motor
proteins that are moving along microtubules.
-invagination would allow membrane adherence
then close off after membrane fusion and then
membrane pocket that is formed will eventually
separate/break off/ pinch off from the PM
WHAT WOULD HAPPEN TO THE VESICLES AND
ITS CONTENTS?
-fused to the lysosomes
-digested-because lysosomes are rich in
hydrolytic enzymesis
if it contains a protein-it will be
hydrolyzed to produce amino acids and
released into the cytosol to be used by
the cell
- fused with endosomes for further processing
-avoid the lysosomes and avoid
hydrolysis,
-transcellular transport/ transcytosis
3 TYPES OF ENDOCYTOSIS
1 Phagocytosis-cell eating
-one cell engulfing another whole cell/very
large structure
-e.g Amoeba engulfing 1 whole yeast cell
 Phagocyte engulf/internalize a
phatogen/bacterial cell inte
-try to internalize something as big as it is
--involve microfilaments in the membrane and
has toadjust the microfilament network
supporting the membrane
microfilament network supporting the
membrane are very important during the
formation of phagosomes
1 Pinocytosis-cell drinking
-what is engulf may be essentially just
extracellular fluid that may just contain some
dissolve substances
-microfilament network is not involve anymore
- vesicles are much smaller, not much
strain on the membrane
1 Receptor-mediated endocytosis
-involve receptors making it more specific,
-makes endocytic processes more
concentrated
In the extracellular fluid there are several
types of molecules,
if this is just a regular endocytosis ,
it's possible that the vesicle contain
heterogeneous mix of molecules
but if you have a receptor,
it will just recognize and binds its
substrates/ligand , so even the ECF
will not be allowed to be part of what
has been taken in (or very small
amount would be included)
-RME, apart from involving receptors,w ould
also COAT proteins, particularly Clathrin-coat
Clathrin Triskelia- monomer of clathrin
coat
-each one is triskelion=three-legged
How they are formed?
On the cytosolic side of the
membrane, a number of
Clathrin Triskelia will now start
to aggregate or group together
to form a pentagonal/hexagonal
structure, the force of their
aggregation causes the
membrane to invaginate and
initiate the formation of coated
vesicles.
-vesicle coat is coated with
proteins
Receptors and Clathrin
e.g On one side of the membrane, they
would recognize and bind their
ligand/cargo, then that cargo-receptor
complex will now move to another area of
the membranes(remember, membrane
proteins can laterally diffuse) , we have
here clathrin aggregating on the cytosolic
side, therefor causing the invagination of
the membrane forming the coated pits
.This receptor-cargo complex will now
laterally diffuse and move into that
coated pit. Then, formation of coated
vesicles, however, the coat will eventually
disassociate itself since they are not
permanently bound to the vesicles, the
clathrin will back and start aggregating
again to form other coated pits, to
participate in another round of RME.
What happens to the uncoated vesicles?
-fuse with lysosomes
-fuse with endosomes
-be involve in transcytosis
E.g UPTAKE OF CHOLESTEROL
Cholesterol is transported in the blood as
part of a complex, as LOW DENSITY
LIPOPROTEIN (LDL) , complex with
proteins and other lipids. Now, in our cell
membrane, we have LDL receptors, so
that receptor recognizes the LDL, and the
LDL-receptor complex moves into a
clathrin-coated pit, initiating a formation
of a coated clathrinvesicle but eventually
the clathrin will be released and recycle
back to the membrane. Now you will only
have a vesicle containing LDL+receptor
and this would fuse with the endosome
(has acidic pH). During the binding of
LDL-receptor with LDL (pH 7=having high to be released but not the
affinity with each other), here inside the apotransferrin (apotransferrin
endosome, pH dropped to 5 and becomes remains bound to the receptor)
acidic, binding affinity decrease between Iron is released into the cytosol
the receptors and the ligands. So receptor and used by the cell
releases the LDL, receptors are recycled Inside the vesicle, receptor is
back to the membrane to be reused. Left bound to its ligand and go back
is endosome containing LDL, no reecptors (exocytosis) exposing the
anymore. Next step, this will fuse with receptor and apotransferrin to
the lysosomes, so there will now be pH 7 and that will decrease
hydrolysis of the LDL, protein components their binding affinity so that the
hydrolyze to amino acids, lipids apotransferrin is released and
components hydrolyze to fatty acids, will pick up more iron.
cholesterol will now be released by At PH 7,receptor binds to
hydrolysis. All will now be release into ferrotransferrin, releasing
the cytosol to be used by the cell. apotransferrin
Take note of the significance of pH At acidic pH, only iron is released.
difference=influence the binding of the
receptor with its ligand. 3. TRANSCELLULAR TRANSPORT/TRANSCYTOSIS of maternal
Any of these processes, will affect the antidbodies (antibodies from breastmilk
cholesterol uptake. How they are transported from the gut into the blood
If mutation would prevent stream of an infant
recognition of LDL, or binding to the Take note of role of change in pH:
coated pit, it will disrupt uptake. In gut-acidic?
In ECF-neutral?
E.G RME of Iron
-Iron is transported bounded to a
protein that is part of
ferrotransferate.
-Protein is apotransfrerin, if there is
iron -ferrotransferrin
Receptor recognizes transferin,
pH 7 high binding affinity between
the transferrin receptor and
ferrotransferrin
Formation of clathrin coated
vesicles
Removal of the clathrin to be
recycled
Vesicle containing the receptor
with ferrotransferrin but this
avoids the lysosome
Fuse with the endosome and
shifts pH to acidic causing iron