CHLOROPLAST

 
2005-2006
 Energy  Conversion  
• Mitochondria convert the chemical energy
of reduced carbon compounds into ATP.
• Chloroplasts convert light energy to the
energy of reduced carbon compounds
• Fundamental to both processes is an
electron transport chain, where energy is
passed from compound to compound in
coupled oxidation:reduction reactions
 General  concepts  
• AFW  Schimper  -­‐-­‐-­‐-­‐  Used  the  term  ‘Plas>d’  
• A  Meyer  -­‐-­‐-­‐-­‐  detailed  cytological  studies  
• Plas>dome  -­‐-­‐-­‐-­‐  gene>c  informa>on  carried  by  
plas>d  
• Plas>d  inheritance  -­‐-­‐-­‐  independent  of  nucleus  
-­‐-­‐-­‐-­‐-­‐  follows  maternal  inheritance  
 Chloroplasts
• Chloroplasts are plant organelles
– class of plant structures = plastids
• leucoplasts
– store starch in roots & tubers
– Amyloplast (starch), Elaioplast (oil), Aleuroplast
(protein)
• chromoplasts
– store pigments for fruits & flowers
• chloroplasts
– store chlorophyll & function
in photosynthesis
– in leaves, other green
structures of plants &
in eukaryotic algae
2005-2006
Plastid development is plastic & mostly under
nuclear control.

Shoots:

proplastids etioplasts chloroplasts

chromoplasts

Roots:
proplastids amyloplasts
 Chloroplasts
• Structure
– 2 membranes
• outer membrane
• inner membrane
– internal fluid-filled space =
stroma
• DNA, ribosomes & enzymes
• thylakoids = membranous sacs where ATP is made
• grana = stacks of thylakoids

Why  internal  sac  membranes?  

increase  surface  area  for    
membrane-­‐bound  enzymes  that  
synthesize  ATP   2005-2006
Chloroplast   Thylakoid membrane –
proteins of ETC, ATP
synthase
• Inner membrane –
membrane transport
protein for Rubisco
• Outer membrane -
porin
• Stroma – enzymes for
Calvin Cycle,
chloroplast DNA.
Transport  of  Proteins  into  Mitochondria  and  Chloroplasts  

•Organelles  specialized  for  ATP  synthesis.  

•Most,  but  not  all,  proteins  are  encoded  by  the  nuclear  genome  and  
synthesized  in  the  cytosol.  
•Proteins  must  be  transported  to  one  of  mul>ple  membranes  or  
compartments.  
Transport into chloroplasts is similar to mitochondria except
there is a 3rd membrane that can be targeted. Targeting the
thylakoid membrane involves a second signal sequence.

In the case of chloroplasts, the electrochemical proton gradient

is at the thylakoid membrane where this gradient participates
in transport. Transport across the chloroplast inner membrane
is powered by GTP and ATP hydrolysis.    
 Endosymbiosis theory
• Mitochondria & chloroplasts were once
free living bacteria
– engulfed by ancestral eukaryote
• Endosymbiont
– cell that lives within another cell (host)
• as a partnership
• evolutionary advantage
for both
– one supplies energy
– the other supplies raw materials
& protection
2005-2006
The Endosymbiotic Theory
 Chloroplast DNA (cpDNA)

General features:
1. double-stranded, circular molecule
2. no histones, but have bound proteins (e.g.,
Hu), organized into nucleoids
3. G-C content typically less than nuclear DNA
4. multiple copies (~30-100) per plastid (i.e., all
cp genes are multi-copy)
5. can be 10-20% of the total DNA in leaves
 Chloroplasts
• Function
– photosynthesis
– generate ATP & synthesize sugars
• transform solar energy into chemical energy
• produce sugars from CO2 & H2O
• Semi-autonomous
• moving, changing shape & dividing
• can reproduce by pinching in two

Who  else  divides  like  

that?  
bacteria!   2005-2006
 Photosynthesis  
• Photosynthesis  occurs  in  thylakoid  membranes  of  
chloroplasts  -­‐  structures  involving  paired  folds  
(lamellae)  that  stack  to  form  "grana"    
• The  soluble  por>on  of  the  chloroplast  is  the  
"stroma"    
• The  interior  of  the  thylakoid  vesicles  is  the  
"thylakoid  space"  or  "thylakoid  lumen"    
• Chloroplasts  possess  DNA,  RNA  and  ribosomes    
 Photosynthesis  Consists  of  Both  
Light  Reac>ons  and  Dark  Reac>ons  

• The  light  reac>ons  capture  light  energy  and  

convert  it  to  chemical  energy  in  the  form  of  
reducing  poten>al  (NADPH)  and  ATP  with  
evolu>on  of  oxygen,  occur  in  grana    
• The  dark  reac>ons  use  NADPH  and  ATP  to  drive  
the  endergonic  process  of  hexose  sugar  forma>on  
from  CO2  in  a  series  of  reac>ons  in  the  stroma    
 Chlorophyll  

Photoreac)ve,  isoprene-­‐based  pigment    

• A  planar,  conjugated  ring  system  -­‐  similar  
to  porphyrins    
• Mg  in  place  of  iron  in  the  center    
• Long  chain  phytol  group  confers  membrane  
solubility    
• an  efficient  absorber  of  light    
 Mitochondria & chloroplasts are different
• Organelles not part of endomembrane system
• Grow & reproduce
– semi-autonomous organelles
• Proteins primarily from free ribosomes in cytosol
& a few from their own ribosomes
• Own circular chromosome
– directs synthesis of proteins produced by own
internal ribosomes

Who  else  has  a  circular  chromosome  no  bound  

within  a  nucleus?  
bacteria   2005-2006
RIBOSOME  
The ribosome is composed of a large and small subunit
 Ribosomes  
• Protein  biosynthe>c  
machinery  
• Made  of  2  subunits  (bacterial  
30S  and  50S,  Eukaryotes  40S  
and  60S)  
• Intact  ribosome  referred  to  as  
70S  ribosome  in  Prokaryotes  
and  80S  ribosome  in  
Eukaryotes  
• In  bacteria,  20,000  ribosomes  
per  cell,  20%  of  cell's  mass.  
• Mass  of  ribosomes  is  roughly  
2/3  RNA  
 ProkaryoKc  Ribosome  Structure  

• E.  coli  ribosome  is  25  nm  

diameter,  2520  kD  in  mass,  and  
consists  of  two  unequal  subunits  
that  dissociate  at  <  1mM  Mg2+    
• 30S  subunit  is  930  kD  with  21  
proteins  and  a  16S  rRNA    
• 50S  subunit  is  1590  kD  with  31  
proteins  and  two  rRNAs:  23S  
rRNA  and  5S  rRNA    
 EukaryoKc  Ribosome  Structure  
• Mitochondrial  and  chloroplast  
ribosomes  are  quite  similar  to  
prokaryoKc  ribosomes,  reflecKng  
their  supposed  prokaryoKc  origin    
• Cytoplasmic  ribosomes  are  larger  and  
more  complex,  but  many  of  the  
structural  and  funcKonal  properKes  
are  similar  
• 40S  subunit  contains  30  proteins  and  
18S  RNA.  
• 60S  subunit  contains  40  proteins  and  
3  rRNAs.  
 Ribosome  Structure  
• Crystal  structure  of  ribosome  is  
known  
• mRNA  is  associated  with  the  30S  
subunit  
• Two  tRNA  binding  sites  (P  and  A  
sites)  are  located  in  the  cavity  
formed  by  the  associaKon  of  the  
2  subunits.  
• The  growing  pepKde  chain  
threads  through  a  “tunnel”  that  
passes  through  the  40S  (30S  in  
bacteria)  subunit.  
 Ribosomal  Proteins    
• L12  -­‐-­‐-­‐-­‐  larger  subunit  protein,  does  not  interact  
with  RNA  
• Four  L7/L12  plus  L10  makes  "L8"    
• Only  one  protein  is  common  to  large  and  small  
subunits:  S20  =  L26    
• Ligle  known  of  structures  -­‐  these  proteins  are  
insoluble  and  difficult  to  study    
The ribosome recycling factor mimics a tRNA
To participate in a new round of polypeptide synthesis, the ri
bosome gets separated from the tRNA and the mRNA and di
ssociate into its large and small subunits in a process called r
ibosome recycling.
RRF (ribosome recycling factor)
The Ribosome is a ribozyme.
The peptidyl transferase center is composed by the 23S rRNA of the lar
ge subunit.
The ribosome has three binding sites for tRNA
The A site is for the aminoacylated-tRNA.
The P site is for the peptidyl-tRNA.
The E site is for the exiting tRNA.
Each tRNA binding site is at the interface between the large and the small
subunits of the ribosome.