CH1131 Biomolecular

Engineering

Week3(Aug25and28)
Organelles,CellGrowthandCytoskeleton

1
LearningObjectives
Organelles
CellGrowth
Cytoskeleton

2
PlasmaMembrane Structure

Lipidbilayer(includingcholesterol)
Glycoproteins
Transmembranestructuralproteins
Transmembranechannels

3
PlasmaMembrane Functions
Compartmentalization continuous
enclosureforspecificactivities

Barrier protectscellularcomponentsbut
ispermeable

Transport machineryfortwowaytraffic
ofsubstances

Response receptorlinkedsignaling
cascadeinresponsetoexternalstimuli

Interaction intercellularrecognition,cell
substrateadhesion

4
 PlasmaMembrane Composition

Membrane lipidprotein
assemblyheldbynoncovalent
bonds

MembraneLipids polarized
OverallComposition
bilayer(outsidehydrophilic)

MembraneLipids mostly
phospholipids(exceptcholesterol)

PhospholipidsinMembrane 5
 PlasmaMembrane Fluidity

Providescompromisebetweenrigidstructureandfluidliquid
Allowsmovementofmembraneproteinsforspecificactivities
Allowsgrowthofmembranewithnewcomponents
Allowscoordinatedmovementofcells

https://www.youtube.com/watch?v=Qqsf_UJcfBc
https://www.youtube.com/watch?v=jM_xePC70Yo 6
 PlasmaMembrane LipidBilayer

LipidBilayer flexibleandtolerate
changesinshape(movementandcell
division)
http://www.youtube.com/watch?v=9wGjkd
WGWNI Membrane Ruffling
ShapeChangeandMovement

LipidBilayer abletoselfassemble
(liposomeanddelivery)

https://www.youtube.com/watch?v=s5fpsl_77gM
https://www.youtube.com/watch?v=lm-dAvbl330

LiposomeandApplications 7
PlasmaMembrane IntegralMembraneProteins

Transmembranebypenetrateinto
lipidbilayer

Hydrophobicportion(usuallyin
helicalstructure)incontactwithin
bilayer

Hydrophilicportionoutsidebilayer

Mayformchannel(hydrophilic
sidewall)fortransport

8
PlasmaMembrane PeripheralMembraneProteins
Noncovalentlyattachedto
hydrophiliclipidendsorintegral
proteinsends

Thosefoundoncytosolicmembrane
surfacefunctionasmembrane
skeletonorsignalingmolecules

Thosefoundonexternalsurfaceare
partofextracellularmatrix

9
 PlasmaMembrane transport

Lipidbilayerpreventslossof
substancesfromcells

Mechanismofexchangeof
substancesinandoutofcells

Membranetransportbypassive
diffusion

Membranetransportbyactive
energydriven process
TypesofTransport
AcrossMembrane

10
 PlasmaMembrane facilitateddiffusion

Definition:diffusionwhereby
substancebindsspecifically to
membranespanningproteinand
diffusesthroughit

Themembranespanningproteinisa
facilitativetransporter

Thereisnoreleaseofenergy in
facilitateddiffusion(vsactive
transport)

DiffusionofGlucose Bindingispossibleonbothsides of
transporter
11
 PlasmaMembrane activetransport

Ionsgradientsareneededand
maintainedbypumpsinactive
transport
Pumpsareenergydriven
1st,3Na+ bindtotransporterfrom
cytoplasmicsideandtriggersits
phosphorylation
2nd,phosphorylationchanges
transportersconformation,allows
Na+/K+ TransportCycle
exitof3Na+
3rd,2K+ bindtooutsideof
transporter,whichisthen
dephosphorylated,leadingto
releaseof2K+ insidethecell
https://www.youtube.com/watch?v=yz7EHJFDEJs 12
 IntracellularOrganelles: mitochondria
Centreofoxidativemetabolism

OxidativemetabolismincorporatesO2 into
metabolismtooxidizecellularcompounds
intoCO2 andH2O

Oxidativemetabolismgeneratesenergy
Mitochondria and Cell neededforcellularactivitiesandstoredas
ATP

13
 IntracellularOrganelles: mitochondrialfunction

MainReactionsinMitochondrion:
glycolysis:degradationofglucosetopyruvatesandgenerates2ATPsfor1
glucose
PyruvateOxidation(Krebscycle):furtherdegradationtoCO2 andH2O,generates
36ATPs
OxidativePhosphorylation:ATPformationcoupledwithenergyreleasedfrom
electrontransport
14
IntracellularOrganelles:
endoplasmicreticulum(ER)

RoughER
ribosomesattached
proteinsecreting
siteofbiosynthesis

SmoothER
lacksribosomes
synthesisofhormone
releaseofglucosefrom
glycogen
sequestingCa2+

15
 PlasmaMembraneandCellular
Organelles summary
Plasmamembraneconsistsoflipidbilayerandproteins(some
arechannels)
Lipidsandproteinscanbelinkedtocarbohydrates(Glycolipids
andGlycoproteins)
Lipidbilayerprovidescellswithflexibilityinmovementand
changingshapes
Proteinscanbeinvolvedinsensingstimuli(receptors)and
movingsubstancesacrossthemembrane(diffusionand
transport)
Mitochondria centreofenergyincells
RoughendoplasmicReticulum proteinsynthesis(comapred
withproteinsynthesisincytoplasm)

16
IntracellularOrganelles:
endomembranesystem
LearningObjectives

Consistsofendoplasmicreticulum,
Golgicomplex,endosomes,
lysosomes,andvacuoles

Transportvesiclestomovematerials
incells

Biosyntheticpathway=secretion

Endocytic pathway=uptake

http://www.youtube.com/watch?v=XXsAf_3
MZNk Golgi and Transport Vesicles 17
IntracellularOrganelles:RoughER
MembraneBiosynthesis
continuedprocessbyfusingwithexisting
membrane
membranecomponentsmovetodifferent
compartments(ER,Golgi)andreceive
modifications
membranecomponentsshowasymmetry
(glycosylation)

SynthesisofMembraneLipids
membranelipidsaremadeinERand
Golgi
phospholipidsaremadebyERmembrane
enzymes
membranelipidsinsertedtoplasma
membranetogetherwithother
components

18
 PlasmaMembrane Carbohydrates
Carbohydrates covalentlylinked
tolipidandproteins

Carbohydrates alwayslocated
outside ofplasmamembrane

CarbohydratesonProtein
(Glycoproteins) functionas
receptors

BloodGroups
CarbohydratesonLipids
Gal:GalactoseGlu:GlucoseFuc:Fucose
(Glycolipids) BloodGroups as
GlcNAc:Nacetylglucosamine
bestknownexample
GlcNAc:Nacetylgalactosamine
19
 IntracellularOrganelles: Golgicomplex

Flattened,disclikemorphology
Golgicomplexispolarized:cis facingER,trans closertoplasma
membrane
Vesiclesbuddingattrans side
Assemblyofglycoproteinsandglycolipids

https://www.youtube.com/watch?v=bD4z27ASN1M 20
 IntracellularOrganelles: vesicletransport
Transportvesiclecoveredwithprotein
coat

Proteincoatsselectcomponentsfor
transport

Threetypesofproteincoats(COP)

COPIIcoatedvesicles:ERtoGolgi

COPIcoatedvesicles:GolgitoER

Clathrincoatedvesicles:trans Golgito
endosome/lysosome,plasmamembraneto
endosome
ER Golgi Intermediate
Compartment
https://www.youtube.com/watch?v=rvfvRgk0MfA21
 IntracellularOrganelles: endosomes

Uptakeofmoleculestoolargefor
transportthroughplasma
membrane

Twomainmechanisms:
phagocytosis:uptakeofpathogens
(bacteria)
endocytosis: uptakeofprotein,
hormones(growthfactors)

Receptor-mediated endocytosis

22
IntracellularOrganelles: endosomes
Phagocytosis(celleating)
phagocyticcellsinclude
macrophages/neutrophils

helptoremovedamaged/dying
cells

phagocytosedmaterialsdigested
bylysosomalenzymes

http://www.youtube.com/watch?v=DuDm
vlbpjHQ Endocytosis and Exocytosis

23
IntracellularOrganelles: endosomes
Endocytosis(receptormediated)

Selectiveandefficientuptakeof
macromoleculesinlowextracellular
concentration

Receptorbindingatplasmamembrane
withclathrincoatatcytoplasmicside

Recyclingofhousekeepingreceptors
(cholesterol)toplasmamembrane,and
cholesterolusedinnewmembrane
biosynthesisandmetabolicprocesses

Degradationofsignalingreceptors(growth
hormones)afteractivationofsignaling
cascade

24
IntracellularOrganelles: lysosomes
Lysosomalenzymesaremadein
ERandmovedtoGolgi

Mannosephosphorylation
distinguisheslysosomalenzymes
fromothers

Lysosomalenzymesare
transportedinclathrincoated
vesicles

25
Intracellular Organelles: lysosomes
Targeting of Lysosomal Enzymes
1. Lysosomal enzymes synthesized in RER

2. Lysosomal enzymes are glycosylated (mannose)

and phosphorylated (in CGN), to form mannose-6-
phosphate side chain

3. Modified lysosomal enzymes are recognised by

mannose-6-phosphate receptors (MPR), with a coat
protein and adaptor protein (), at TGN and form
vesicles

4. Coat protein and adaptor dissociate from vesicle,

lysosomal enzymes delivered to endosome and
vesicle with MPR

5. MPR returned to TGN to start the cycle

6. MPR can also be present at plasma membrane, to

capture secreted lysosomal enzymes to direct them
to lysosome 26
 IntracellularOrganelles: proteasome

Largemultiplecatalyticcomplex
localizedbothinthenucleusandin
thecytoplasm(associatedwithER)

Intracellularorganellesfor
degradationofmisfoldedproteins

Intracellularorganellesfor
processingofpeptideforantigen
presentation

http://www.youtube.com/watch?v=4DMqnfrzpKg Proteasome

27
 IntracellularOrganelles: summary

GolgiComplex transport
Endosome uptakeoflargemolecules
Lysosomes digestivesystemofcells
Proteasome degradationofmisfoldedprotein

28
CytoskeletonandMotility: learningobjectives

Describetraitsandfunctionsof
variouscytoskeletonelements

Describestructuresandfunctionsof
molecularmotors

Describestructuresandfunctionsof
microtubules

29
 Cytoskeleton: overviewofmajorfunctions

Components:
Microtubules,(IntermediateFilaments)andMicrofilaments

Functions:
Scaffold,HoldingOrganelles,IntracellularTransport,Movement,CellCycle
30
Microtubules:structureandfunction
Hollow,tubularstructurewithprotofilaments
arrangedinlongitudinalrows

Eachprotofilamentisassembledfromdimeric
buildingblocks( andtubulin)

Microtubulesdisplaypolarity:
plusendhasantubulinandisfastgrowing
minusendhasantubulinandisslowgrowing

MicrotubtulesarestabilizedbyMicrotubule
AssociatedProteins(MAPs)

31
 Microtubules: tubulin

tubulinhasaboundGTPwhichisnot
hydrolyzed

tubulinhasaboundGDPwhichcanbe
exchangedforGTP

https://www.youtube.com/watch?v=21P100yiEN8
Structure of tubulin http://www.youtube.com/watch?v=PvDlilBgoSs
Microtubule Assembly

32
 Microtubules: functions
Structuralsupport (polesintent)

Passive structuresandserveastrack
formotorproteinsinintracellular
transport
forexample,providesintracellular
Motilityinelongatedcellularprocesses
(neuronalcells)

Cellular Localization of Tubulin Maintain IntracellularOrganellesin

place

Cytoplasmicmicrotubulesdetermine
cellshape

MT in Nerve Cell
33
 Microtubules: MotorProteins
MotorProteinsconvertchemicalenergy (ATP)into
mechanicalenergy forcellulartransport

Materialsintransportincludevesicles,organelles,
chromosomesandcytoskeletalelements

Threemainfamiliesofmotorproteins:Myosins (for
microfilaments),Kinesins (formicrotubules),Dyneins (for
microtubules)

HydrolysisofATP releasesmechanicalenergyfor
movementofmotorproteins

http://www.youtube.com/watch?v=7sRZy9PgPvg Motor Proteins

34
 Microtubules: KinesinsasMotorProteins
Movesvesicles/organellesinnerve
cells
Composedoftetramer(2heavyand2
lightchains)
Headsbindtomicrotubulesand
generatemechanicalenergy
Structure of Kinesin Tailsbindtotransportingmaterials
Kinesinmovementisprocessive
2headsareindifferentstagesof
chemicalandmechanicalcycle
Conformationchangein1headafter
ATPbindingmovestheotherhead
Twomechanismsofkinesin
movement
Movement of Kinesin
35
 Microtubules: DyneinasMotorProteins
Mainfunctionintransporting
chromosomeduringmitosisand
Golgi/otherorganelles
Transportinoppositedirection
comparedwithkinesins
Largeprotein(1.5X106 Da)
Composedof2heavychainsand
varietyofintermediate/light
Structure of Dynein chains
Headsactsasenergygenerating
site
Stalksinvolvedinbindingto
microtubules
Lightchainsinvolvedinbinding
transportingmaterials
36
 Microtubules: DyneinversusKinesins

Dyneinmovestowardsminusendofmicrotubules
Kinesinsmovetowardsplusendofmicrotubules
37
 Microtubules: dynamicproperties

Changing Distribution of Microtubules during Cell Division

Cytoplasmicmicrotubulesaredynamic(assembleanddisassembly)
Mechanismsformicrotubulereorganisation:
rearrangementofexistingmicrotubules
disassemblyofexistingmicrotubulesandreassemblyofnewones
dynamicchangesassociatedwithGTPhydrolysis
38
 Microfilaments:structuresandfunctions
Microfilament(MF),actinfilamentand
Factinaresynonyms

Actinpresentandhighlyconservedin
sequenceinalleukaryoticcells

Actinkeyelementincellmotileprocess

MonomerGactinpolymerizestoform
filamentmadeof2strandsofpolymer

Actinfilamentdisplayspolarity

Morphology of Actin
39
 Microfilaments: assemblyanddisassembly
ActinmonomerbindsATPbefore
polymerization

ActinabletohydrolyzeATP

Actinpolymerizationoccursfasterat
plusend

Cellmaintainsdynamicbalance
betweenassemblyanddisassembly
ofactin

Dynamicbalancerequiredforcell
movement,shapechangeandcell
division
Actin Assembly

http://www.youtube.com/watch?v=Yt59kx_z6xM Actin Polymerization 40

 Microfilaments: myosinasmotorprotein
Solefunctionasmotorforactin
Conventional myosin(typeII)
foundinmostmuscleandnon
musclecells
HeadbindtoactinandATP
(distinctdomains)
2lightchainsareassociatedand
formfilaments

Structure of Myosin

41
 RolesofActin
Cellmovementcommoninmanyprocesses
(develoment,woundhealing,tumourcellspreading)

Cellmovementdependsondynamicbalance
ofactinassemblyanddisassembly

Movement of Cultured Cell Vesicletrafficking

Lateralmovementofmembraneproteins

Cytokinesis

Phagocytosis

42
 CellMovement:signalingpathway
Actinpolymerizationrequiredfor
cellmovementandchangein
shape
Cellshapechangemediatedby
signalingmolecules
Signalingmoleculesdisplaysbinary
Cell Shape Change and Movement switchbystateofGTP
Onstate:GTPboundandsignaling
activated
Offstate:GDPboundandsignaling
shutdown

Regulation of Rho p21s 43

 CytoskeletonandMotility: summary
CytoskeletonconsistsofMicrotubules,(IntermediateFilaments)and
Microfilaments
CytoskeletonsmainfunctionincludeScaffold,HoldingOrganelles,
IntracellularTransport,MovementandCellCycle
Microtubules(MTs)consistofdimericbuildingblocks( andtubulin),
bindGTPandprovidestructuralsupporttocellsandtrackformotor
proteins(kinesinsanddynein)
Microfilament(MF)oractinfilamentbindsATPandisakeyelementincell
motileprocess,withmyosinasmotorprotein
Actinpolymerizationanddepolymerization,criticalforcellmovement,is
regulatedbyintracellularsignalingpathway

44
 TutorialTopics

Locationofreceptors(otherthanplasma
membrane)?

Howdoescellmaintainthesizeoforganelles?