Transport of Systems in Microbial

Cells
Presented by-
Diptomoy Bhattacharya

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 Transport across cell membrane
 All cells are generally separated from their surrounding environment by
plasma membrane.
 In addition, the eukaryotic cells are compartmentalized by intracellula
r membranes that form the boundaries and internal structures of variou
s organelles.
 These biological membranes are semi-permeable in nature that is their p
ermeability properties ensure that the specific molecules and ions read
ily enter the cell and the waste products leave the cell.
 These movements of solutes into the cell are mediated through the actio
n of specific transport proteins that are present on the cell membrane.
Such proteins are therefore required for movements of ions, metabolites
and even water.
 Transport proteins are also responsible for biological electrochemical
phenomena such as neurotransmission.

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 Cell membrane architecture in transp
ort across cell membrane:
 The cell membrane plays an important role in transport of molecul
es. Because it acts as a semi-permeable barrier, allowing specifi
c molecules to cross while fencing the majority of organically pr
oduced chemicals inside the cell.
 Electron microscopic examinations of cell membranes reveal the de
velopment of the lipid bilayer model (fluid-mosaic model).The mod
el consists ofphospholipid, which has a polar (hydrophilic) head
and two nonpolar (hydrophobic) tails.
 These phospholipids are aligned tail to tail so the non-polar ar
eas form a hydrophobic region between the hydrophilic heads on th
e inner and outer surfaces of the membrane

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 Permeability of molecules across pho
spholipid bilayer:
 Most of the molecule will diffuse across a protein-free lipid bilayer do
wn its concentration gradient, if provided enough time.
 The diffusion rate is the function of the size of the molecule and its r
elative solubility in oil. In general, the smaller the molecule and the
more soluble in oil (the more hydrophobic or non-polar), the more rapidl
y it will diffuse across a cell membrane.
 Small non-polar molecules, such as O2 and CO2, readily dissolve in cell
membrane and therefore diffuse rapidly across them whereas small uncharg
ed polarmolecules, such as water or urea, also diffuse across a bilayer,
but much more slowly but ethanol diffuses readily.

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 • Conclusively it can be said that lipid bilayers are highly impermeable to charged molecu
les (ions) by considering its size also because the charge and high degree of hydration
of such molecules prevents them from entering the hydrocarbon phase of the bilayer. Thus
, these bilayers are 109 times more permeable to water than to even such small ions as N
a+ or K+

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 Thermodynamics of transport :
 The diffusion of a substance A, across the two sides of a membrane
thermodynamically resembles a chemical equilibration.
 A(out) → A(in)
 In the following sections, the free energy of a soluteA, varies wit
h its concentration:

 where GA is the chemical potential (partial molar free energy) of A

(the bar indicates quantity per mole) and G°`A is the chemical pote
ntial of its standard state.
 Thus, a difference arises in the concentrations of the substance
on two sides of a membrane and generates a chemical potential diffe
rence:

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 If the concentration of A outside the membrane is greater than that insid
e, ΔGA for the transfer of A from outside to inside will be negative and
the spontaneous net flow of A will be inward. Conversely, if [A] is great
er inside than outside, ΔGA is positive and an inward net flow of A can
occur only if an exergonic process, such as ATP hydrolysis, is coupled to
it to make the overall free energy change.The transmembrane movement of i
ons also depends in charge differences across the membrane, thereby gener
ating an electrical potential difference which is given by:
 ΔA=A(in) – A(out),
 whereΔAis termed the membrane potential.Consequently, if A is ionic, mus
t be amended to include the electricalwork required to transfer a mole of
A across the membrane from outside to inside:

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 Non-mediated transport
 Non-mediated transport occurs through the simple diffusion proce
ss and the driving force for the transport of a substance throug
h a medium depends on its chemical potential gradient.

Mediated transport
 Whereas mediated transport requires specific carrier proteins. T
hus, the substance diffuses in the direction that eliminates its
concentration gradient; at a rate proportional to the magnitude
of this gradient and also depends on its solubility in the membr
ane’s non-polar core.

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 Mediated transport is classified into two categories depending on the the
rmodynamics of the system:
 1. Passive-mediated transport, or facilitated diffusion:In this typeof pr
ocess a specific molecule flows from high concentration to low concentrat
ion.
 2. Active transport:In this typeof process a specific molecule is transpo
rted from low concentration to high concentration, that is, against its c
oncentration gradient. Such an endergonic process must be coupled to a su
fficiently exergonic process to make it favorable (ΔG <0).

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 Passive mediated transport:
 Substances that are too large or polar diffuse across the lipid b
ilayer on their own through membrane proteins called carriers, pe
rmeases, channels and transporters. Unlike active transport, this
process does not involve chemical energy. So the passive mediated
transport is totally dependent upon the permeability nature of ce
ll membrane, which in turn, is function of organization and chara
cteristics of membrane lipids and proteins.
 Passive or simple diffusion allows for the passage across the cel
l membrane of simple molecules and gases, such as CO2, O2, and H2
O. In this case, a concentration gradient must exist, where there
is higher concentration of the substance outside of the cell than
there is inside the cell. As more of the substance is transported
into the cell the concentration gradient decreases, slowing the r
ate of diffusion.

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 Types of Passive transport
 Diffusion:
 The process of the net movement of solutes from a region of high
concentration to a region of low concentration is known as diffu
sion. The differences of concentration between the two regions a
re termed as concentration gradient and the diffusion continues
till the gradient has been vanished. Diffusion occurs down the c
oncentration gradient.

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 Facilitated diffusion :
 The process of the movement of molecules across the cell membrane via spe
cial transport proteins that are embedded within the cellular membrane is
known as facilitated diffusion or called carrier-mediated diffusion. Many
large molecules, such as glucose, are insoluble in lipids and too large t
o fit into the porins, therefore, it will bind with its specific carrier
proteins, and the complex will then be bonded to a receptor site and move
d through the cellular membrane.
 Facilitated diffusion also involves the use of a concentration gradient,
where the concentration of the substance is higher outside the cell, but
differs with the use of carrier proteins (sometimes called permeases). Th
ese proteins are embedded within the cell membrane and provide a channel
or pore across the membrane barrier, allowing for the passage of larger m
olecules. If the concentration gradient dissipates, the passage of molecu
les into the cell stops. Each carrier protein typically exhibits specific
ity, only transporting in a particular type of molecule or closely relate
d molecules.

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 Filtration :
 Filtration is the process of the movement of water and solute molecules acr
oss the cell membrane due to hydrostatic pressure generated by the system.
Depending on the size of the membrane pores, only solutes of a certain size
may pass through it.

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 Osmosis
 Osmosis is the type of diffusion of water molecules across a semi- permeable
membrane, from a solution of high water potential to a region of low water po
tential. A cell with a less negative water potential will draw in water but t
his depends on other factors as well such as solute potential (pressure in th
e cell e.g. solute molecules) and pressure potential (external pressure e.g.
cell wall).

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Active transport:
 Active transport is the movement of a substance against its concentration g
radient (i.e. from low to high concentration). It is an endergonic process
that, in most cases, is coupled to the hydrolysis of ATP.
.
Types of Active transport:
1)Primary Active transport: Primary active transport,
also called direct active transport, directly uses energy
to transport molecules
across a membrane.

Example:Sodium-potassium pump,
which helps to maintain the cell potential

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 • Primary active transport involves the use of chemical energy, such as A
TP, to drive the transport. One example is the ABC system, which utiliz
es ATP-Binding Cassette transporters. Each ABC transporter is composed
of three different components:
1) membrane-spanning proteins that form a pore across the cell membrane
(i.e. carrier protein),
2) an ATP binding region that hydrolyzes ATP, providing the energy for t
he passage across the membrane
3) a substrate-binding protein, a peripheral protein that binds to the a
ppropriate substance to be transporter and ferries it to the membrane-
spanning proteins.
 In gram negative bacteria the substrate-binding protein is located in t
he cell’s periplasm, while in gram positive bacteria the substrate-bin
ding protein is attached to the outside of the cell membrane.

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 Group translocation
 It is a distinct type of active transport, using energy from an energy-
rich organic compound that is not ATP. Group translocation also differs
from both simple transport and ABC transporters in that the substance b
eing transported is chemically modified in the process.
 One of the best studied examples of group translocation is the phosphoe
nolpyruvate: sugar phosphotransferase system (PTS), which uses energy f
rom the high-energy molecule phosphoenolpyruvate (PEP) to transport sug
ars into the cell. A phosphate is transferred from the PEP to the incom
ing sugar during the process of transportation.

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Secondary active transport:
 Secondary active transport or co-transport, also uses energy to t
ransport molecules across a membrane; however, in contrast to pri
mary active transport, there is no direct coupling of ATP; instea
d, the electrochemical potential difference created by pumping io
ns out of the cell is instrumental.
 Secondary active transport utilizes energy from a proton motive f
orce (PMF). A PMF is an ion gradient that develops when the cell
transports electrons during energy-conserving processes. Positive
ly charged protons accumulate along the outside of the negatively
charged cell, creating a proton gradient between the outside of t
he cell and the inside.

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 There are three different types of transport events for simple transport:
 uniport, symport, and antiport and each mechanism utilizes a different p
rotein porter. 

 Uniporters transport a single substance across the membrane, either in or

out. 
 Symporters transport two substances across the membrane at the same time,
typically a proton paired with another molecule.
  Antiporters transport two substances across the membrane as well, but in
opposite directions. As one substance enters the cell, the other substanc
e is transported out

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 Examples:
 (Na+–K+)–ATPase
 (Na+ - K+)–ATPaseactive transport system is commonly found in the plasma
membranes of higher eukaryotes, This transmembrane protein consists of tw
o types of subunits: a 110-kD non-glycosylated α- subunit that contains
the enzyme’s catalytic activity and ion-binding sites, and a 55-kD glyco
protein β-subunit of unknown function. Sequence analysis suggests that
the α- subunit has eight transmembrane α-helical segments and two large
cytoplasmic domains. The β- subunit has a single transmembrane helix and
a large extracellular domain. The protein may function as an (αβ)2 tetr
amer in vivo.
 The (Na+ – K+)–ATPase is also called as the (Na+ – K+) pumpbecause it
pumps 3 Na+ out of and 2 K+ into the cell in presence of hydrolysis of in
tracellular ATP. The overall stoichiometry of the reaction is:
 3 Na+(in) + 2 K+(out) + ATP + H2O →3 Na+(out) + 2 K+(in) + ADP + Pi

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 Difference between Active and Pas
sive Transport
Features Primary active transport Secondary active
transport

Other name: Direct active transport Coupled transport or

cotransport

Coupling of ATP Direct coupling of ATP No direct coupling of

ATP

Energy used: Metabolic energy (ATP) Electrochemical gradient

Redox energy
Photon energy

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Features Primary active transport Secondary active
transport
Membrane protein •P-type ATPase, e.g. sodium potassium •Cotransporters:
transporter (ion pump, calcium pump, proton pump •Antiporters
pumps, ion •F-ATPase, e.g. mitochondrial ATP •Symporters
channels, ATPases): synthase, chloroplast ATP synthase
•V-ATPase, i.e. vacuolar ATPase
•ATP-binding cassette transporter: e.g.
MDR, CFTR
Examples: •Active transport using ATP via sodium- •Active transport of a
potassium pump to move 3 Na+ ions out second substrate while
while moving 2 K+ ions into the cell another ion, typically
•Active transport using Redox energy (of Na+, K+, or H+ ions, move
NADH) to generate a proton gradient in down the concentration
the inner mitochondrial membrane gradient
•Active transport using photon energy
(light) to generate a proton gradient
during photosynthesis

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 Iron Uptake
 Iron is required by microbes for the function of their cytochromes and e
nzymes, resulting in it being a growth-limiting micronutrient. However,
little free iron is available in environments, due to its insolubility.
Many bacteria have evolved siderophores, organic molecules that chelate
or bind ferric iron with high affinity. Siderophores are released by the
organism to the surrounding environment, whereby they bind any available
ferric iron. The iron-siderophore complex is then bound by a specific re
ceptor on the outside of the cell, allowing the iron to be transported i
nto the cell.

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 THANK
YOU
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