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DEPARTMENTAL PRESENTATION
DR. SUJIT KUMAR
BSc.(BIOTECH), MD(PHYSIOLOGY)
CCH, CGO, SVD (PUNE)
SCMT KANPUR
PRESENTED BY
GUIDED BY
DR. RYE GHOSE PhD.
SCMT, KANPUR
6 Structure/Function of the Cytosol
Cytoskeleton and Cell Motility (2)
Microtubule-Organizing Centers – (MTOCs)
Assembly – two phases
Nucleation
Elongation
Best studied – Centrosome
Two barrel shaped centrioles
Pericentriolar material (PCM)
Sites where microtubules converge
Cytoskeleton
Microtubule-Organizing Centers – (MTOCs)
Centrioles
PCM – initiates formation of microtubules
Microtubule minus end in centriole
Microtubules enlongated at opposite end
Basal Bodies and other MTOCs
Cilia & Flagella
Identical in structure to centrioles
Cytoskeleton
Microtubule-Organizing Centers – (MTOCs)
Microtubule nucleation
All MTOCs – a common protein – γ-tubulin
Cytoskeleton
Microtubules Dynamic Properties
Mostly extremely labile
Non-covalent bonds
More stable forms
Stabilized by
MAPs
Enzymatic modification
Cytoskeleton
Microtubules Dynamic Properties
Mostly extremely labile
GTP required for assembly
GTP bound to β-tubulin
GTP hydrolysis after incorporation
After dimer is released from structure – GDP replaced
by GTP
A dimer with GTP bound has a different conformation
from a GDP-bound dimer
Cytoskeleton
Microtubules Dynamic Properties
Growing microtubule
+ end is an open sheet
GTP dimers added
Added more rapidly than GTP can be hydrolysed
GTP ‘cap’ favors addition of more dimers
Microtubules can shrink very rapidly
If open end becomes ‘closed’ – the structure becomes
unstable
Cytoskeleton
Microtubules – Cilia and Flagella –
structure/function
Cilia and Flagella – two versions of the same structure
Patterns of movement
Cilia – power stroke – rigid state
- recovery stroke – flexible
Occur in large numbers
Beating is coordinated
Flagella – longer
Different waveform patterns
Cytoskeleton
Microtubules – Cilia and Flagella – structure
Core – axoneme
Microtubule array – 9 peripheral doublets + central pair
+ ends at tip & - ends at base
Each doublet
One complete (13 subunits) - A tubule
One incomplete (10-11 subunits) – B tubule
Cytoskeleton
Microtubules – Cilia and
Flagella – structure
Central tubules
Enclosed by projections -
Central sheath
Connected to A tubules of
peripheral doublets by radial
spokes
Doublets connected to each other
– interdoublet bridge
Interdoublet bridge – an elastic
protein – nexin
Radial spokes in groups of three.
Basal body – A, B and C tubules
Cytoskeleton
Microtubules – Cilia
and Flagella –
structure
Dynein arms
Swinging cross-
bridges
Project from one
doublet
‘Walk’ along the next
So doublets ‘slide’
relative to each other
Cytoskeleton
Intermediate Filaments
Only in animal cells
Interconnected by cross-bridges of plectin
Plectin
Different isoforms
One end –binds IF
Other end varies – isoforms
Another IF
Microtube
Microfiber
Heterogenous group
> 50 genes
6 major classes
Cytoskeleton
Intermediate Filaments
Cytoskeleton
Intermediate Filaments
All classes have
Central, rod-shaped α-
helical domain
Flanked by variable
globular domains
rod-shaped α-helical
domains
Spontaneously form coiled
coils
Both with same polarity
Dimer has polarity
Cytoskeleton
Intermediate
Filaments
Assembly
Tetramer of 2 dimers
Staggered
Antiparallel
Tetramers lack
polarity
Distinguishing
characteristic
Cytoskeleton
Microfilaments
Globular protein –actin
ATP- actin polymerizes
Two strands of actin
Wound around each other
Double helix
Actin filament = F-actin = Microfilament
F-actin – often for in vitro form
Each actin unit has polarity
All actin units in same orientation
Whole filament has polarity
Cytoskeleton
Microfilaments
Arrangement
variable
Highly ordered
arrays
Loose networks
Well defined
bundles
Cytoskeleton
Microfilaments
A major contractile protein of muscle
Occurs in every cell
A major protein
Interacts specifically with myosin
Cytoskeleton
Microfilament – assembly / disassembly
Prior to incorporation
Actin monomer binds to ATP
Actin is an ATP-ase
ATP hydrolyzed after incorporation
During assembly (rapid)
Filament has an actin-ATP cap
Favors assembly
+ end is the fast-growing end
- end
Slow growing
Site of preferential depolymerization
Cytoskeleton
Microfilament – assembly / disassembly
Monomers tend to move down the filament
‘treadmilling’
Intracellular equilibrium – monomeric actin and
polymer
Cellular control of this equilibrium
Localized protein interactive effects
Dynamic reorganization
Locomotion
cytokinesis
Cytoskeleton
Microfilament – assembly / disassembly
Cytoskeleton
Myosin – the molecular motor for Actin
Filaments
Myosin superfamily
Conventional (Type II) myosins
Cytoskeleton
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DR SUJIT 1ST YR MD PHYSIOLOGY cilia & flagella

  • 1. DEPARTMENTAL PRESENTATION DR. SUJIT KUMAR BSc.(BIOTECH), MD(PHYSIOLOGY) CCH, CGO, SVD (PUNE) SCMT KANPUR PRESENTED BY GUIDED BY DR. RYE GHOSE PhD. SCMT, KANPUR
  • 2. 6 Structure/Function of the Cytosol Cytoskeleton and Cell Motility (2)
  • 3. Microtubule-Organizing Centers – (MTOCs) Assembly – two phases Nucleation Elongation Best studied – Centrosome Two barrel shaped centrioles Pericentriolar material (PCM) Sites where microtubules converge Cytoskeleton
  • 4. Microtubule-Organizing Centers – (MTOCs) Centrioles PCM – initiates formation of microtubules Microtubule minus end in centriole Microtubules enlongated at opposite end Basal Bodies and other MTOCs Cilia & Flagella Identical in structure to centrioles Cytoskeleton
  • 5. Microtubule-Organizing Centers – (MTOCs) Microtubule nucleation All MTOCs – a common protein – γ-tubulin Cytoskeleton
  • 6. Microtubules Dynamic Properties Mostly extremely labile Non-covalent bonds More stable forms Stabilized by MAPs Enzymatic modification Cytoskeleton
  • 7. Microtubules Dynamic Properties Mostly extremely labile GTP required for assembly GTP bound to β-tubulin GTP hydrolysis after incorporation After dimer is released from structure – GDP replaced by GTP A dimer with GTP bound has a different conformation from a GDP-bound dimer Cytoskeleton
  • 8. Microtubules Dynamic Properties Growing microtubule + end is an open sheet GTP dimers added Added more rapidly than GTP can be hydrolysed GTP ‘cap’ favors addition of more dimers Microtubules can shrink very rapidly If open end becomes ‘closed’ – the structure becomes unstable Cytoskeleton
  • 9. Microtubules – Cilia and Flagella – structure/function Cilia and Flagella – two versions of the same structure Patterns of movement Cilia – power stroke – rigid state - recovery stroke – flexible Occur in large numbers Beating is coordinated Flagella – longer Different waveform patterns Cytoskeleton
  • 10. Microtubules – Cilia and Flagella – structure Core – axoneme Microtubule array – 9 peripheral doublets + central pair + ends at tip & - ends at base Each doublet One complete (13 subunits) - A tubule One incomplete (10-11 subunits) – B tubule Cytoskeleton
  • 11. Microtubules – Cilia and Flagella – structure Central tubules Enclosed by projections - Central sheath Connected to A tubules of peripheral doublets by radial spokes Doublets connected to each other – interdoublet bridge Interdoublet bridge – an elastic protein – nexin Radial spokes in groups of three. Basal body – A, B and C tubules Cytoskeleton
  • 12. Microtubules – Cilia and Flagella – structure Dynein arms Swinging cross- bridges Project from one doublet ‘Walk’ along the next So doublets ‘slide’ relative to each other Cytoskeleton
  • 13. Intermediate Filaments Only in animal cells Interconnected by cross-bridges of plectin Plectin Different isoforms One end –binds IF Other end varies – isoforms Another IF Microtube Microfiber Heterogenous group > 50 genes 6 major classes Cytoskeleton
  • 15. Intermediate Filaments All classes have Central, rod-shaped α- helical domain Flanked by variable globular domains rod-shaped α-helical domains Spontaneously form coiled coils Both with same polarity Dimer has polarity Cytoskeleton
  • 16. Intermediate Filaments Assembly Tetramer of 2 dimers Staggered Antiparallel Tetramers lack polarity Distinguishing characteristic Cytoskeleton
  • 17. Microfilaments Globular protein –actin ATP- actin polymerizes Two strands of actin Wound around each other Double helix Actin filament = F-actin = Microfilament F-actin – often for in vitro form Each actin unit has polarity All actin units in same orientation Whole filament has polarity Cytoskeleton
  • 19. Microfilaments A major contractile protein of muscle Occurs in every cell A major protein Interacts specifically with myosin Cytoskeleton
  • 20. Microfilament – assembly / disassembly Prior to incorporation Actin monomer binds to ATP Actin is an ATP-ase ATP hydrolyzed after incorporation During assembly (rapid) Filament has an actin-ATP cap Favors assembly + end is the fast-growing end - end Slow growing Site of preferential depolymerization Cytoskeleton
  • 21. Microfilament – assembly / disassembly Monomers tend to move down the filament ‘treadmilling’ Intracellular equilibrium – monomeric actin and polymer Cellular control of this equilibrium Localized protein interactive effects Dynamic reorganization Locomotion cytokinesis Cytoskeleton
  • 22. Microfilament – assembly / disassembly Cytoskeleton
  • 23. Myosin – the molecular motor for Actin Filaments Myosin superfamily Conventional (Type II) myosins Cytoskeleton