:

. , Ph.D.
15-12-2016
1
Y

. H2O2
-

(--)

2
.

1%

: 4-5 ()

3
 E

COS 7 cell expressing an ER marker (red) and a

peroxisomal marker (green).

Bar = 10 microns

movement-2

(Peter Kim) Video 1

(, , ,
)

4
 E

() (C) MIT

(60 )

: , , (pexophagy),

: 1)
2)

5
 (Peroxisomes)..
:
Rhodin, 1954,
Porter & Caulfield, 1958,
De Duve:
22
()

( ) :

O (. )
H2O2,

atrix:
( )

6
 E

() () (Peroxisomes
microbodies)

: (0.1 to 1 m in diameter) (,
)

() 60 80

DNA

: : 1)
FAD H2O2 2) ,

>50
-

7
 (Peroxisomes)..

8
 ()

-
(enz, DAB )
-

/

,
O

10


-
1. peroxisomal membrane proteins (PMPs) ( )

2. Peroxins () ( )
32 peroxins.
matrix

PEX
14 ()


(Peroxisome biogenesis disorders ,
PBDs)

11
 (glyoxysomes)
( )

(glycosomes):
trypanosomes

Woronin-bodies,
filamentous fungi (1
function: plug septal
pores in case of
hyphal injury)

(peroxisomes)

:
, 22 ,
(, )
OIKO

/
(-
- ) 22 12

(. Dictyostelium discoideum)

( Leishmania
() and Trypanosoma spp, ) :

( )

, ( )
(
K )

2 ( )
( )
2 ( )
:
Electron micrograph of Trypansoma brucei 13

Trichomonas ( MIT)

+ CO2 +

e- ( )
2 2: 2
2 ( )
+: 2

( ,
,
)

14


22

/

15


:
-oxidation
H2O2

( )

Woronin bodies

( )

Woronin bodies in a septum of Fusarium oxysporum.

(Wergin, W.P. 1973. Protoplasma 76:249-260
Springer-Verlag). 16


() ()
( )
( , - - )

:
() 2 ()
- -
ROS, RNS

( ) H2O2 (
)

(. )
(oxidation).

17


= 2

: 2 .
2:
:
2

18


,
( )
2 (ATP)
.

E: ,
2

19


:
( , , )




( )

20


Smith and Aitchison, ature

Reviews, Molecular Cell Biology
14:803, Dec 2013

21


,

A =
( ) ester bond ether bond
(vinyl ether phospholipids, plasmenylethalomines,
plasmenylcholines)
:
( )

:
2
( )

PE, PC, PS

ROS

: ,

:
-
22


= : -
- MAVS (mitochondrial antiviral signalling) -

MAVS
-

Smith and Aitchison, ature Reviews, Molecular Cell Biology 14:803, Dec 2013

23


400-1,000 / ()

:
(, ??)

(
)

24

1) 22

10% 2

RH2 + O2 R + H2O2
()
2
(R) 22
22

R:

H2O2 + RH2 22 + R

H2O2 H2O + O2

* H2O2 22

22 -
25


H2O2 + RH2 22 + R peroxidation reaction

22 (. ,
, )

T
25%-50% (EthOH)
.

MetOH, ,

26

2) -

2 + 1/22

CH3OH + O2 HCHO + H2O2

22

CH3OH + H2O2 HCHO + 2H2O

2HCHO + 2NAD + O2 2CO2 + 2NADH

NADH

NADH + ADP + Pi F-ATP NAD + ATP

27


2) -

:
:

(
)
: :

:
acetyl CoA -
28
 :

- (
) &

( )
--
--
-CoA
D-
:


( / )
29

22

3 : (: 170 190 kDa)

(: 225 250 kDa)
(: 240 kDa)

(4 4 )

( = )

30



1) PT-S1R (Pex5p):
& (
KANL COOH-
)

2)

(Pex14p)

31



(vs. MIT, -
)
????

1. .
2. -?

: transient import structures (transient pore hypothesis)

().

Rucktschel et al., Biochimica et Biophysica Acta 1808:892900, 2011)

Erdmann, R. & Schliebs, W. Peroxisomal matrix protein import: the transient pore model. Nature Rev. Mol. Cell
Biol. 6, 738742 (2005).
32
3) -

EN:


(
)
-CoA

33

- /
3) -

- =
2 C
acetyl CoA.
-CoA

( )
( & )

:

- (25-
50%) & (Lazarow & de Duve)

34
3) -

: FADH2
O2 22 ( - H2O 2)
: FADH2 e-

35
3) -


:

- :

(very long chain fatty acids, VLFCA),
C22 ,
(. pristanic acid),

- :
,
(C18
).

36
3) -

1. (10 20 C)
CoA

2.

CoA ()
-CoA ()

-CoA
- *

37
3) -

:
(. - CoA
)
VLCFAs (> 22C) (
)
( C)
MIT

(*** )

38
3) -

- :

3) -

: -
3 (. phytanic acid)
- -
-: phytanoyl-CoA 2-hydroxylase, 2-hydroxyphytanoyl-CoA lyase
pristanal dehydrogenase
1C 2-methyl fatty acid
-

39
4)

alanine-glyoxylate aminotransferase
(AGT)

:
oxalate

.

, oxalate calcium oxalate

( )

40
5) ( )

41
T -

very long chain dicarboxylic

fatty acids acids

ABCD
-

42

Waterham et al., Biochimica et Biophysica Acta 1863:922, 2016



1)
2)

(, )

43


( , ,
)

-, H2O2
( ,
)

44


:


&
Krebs: -CoA 2 CO2
: 2 -CoA (
)

Krebs

5


2/5
45


-
CoA

CoA

Krebs

To

46
 ():

2x

47
 KREBS

:
( / )
( , ) 48


( )

:
2
CO2

49
 :

( )
cyt

50


51


52

:

53

4 :
I. (
)
II. : - -matrix
III.
IV. (),

, Chinese hamster ovary (CHO) :

peroxins ( PEX)

Perxins () ( )

DNA, /

DNA

:
1. (peroxisomal matrix, )
2.

54
H

Schliebs et al., ature Reviews, Molecular Cell Biology 11:885; 2010

55


matrix ,

growth and division

(fission)

56

-
(PMPs)

3 :
(11)
(11)
(DLP1/DRP1, FIS1, Mff GDAP1,
) GTP .

PEX11

11

Waterham et al., Biochimica et Biophysica Acta 1863:922, 2016

57
 DLP1/DRP1 11

Fis1 and Mff

DLP1-/-, Fis1-/- Mff-/-:

DLP1-/-: DLP1

58
 de novo

- (. - PEX3, PEX16
PEX19)

( PMPs -
)
Waterham et al., Biochimica et Biophysica Acta 1863:922, 2016 59
 de novo -
( - all new )
- ,
(PMPs) (
11).

Waterham et al., Biochimica et Biophysica Acta 1863:922, 2016

Smith and Aitchison, ature Reviews, Molecular Cell Biology 14:803, Dec 2013
60


(pexophagy) = .

ubiquitinated
.

(. p62 NBR1)
.

.
Kim P.K ., Hailey D.W., Mullen R.T., Lippincott-Schwartz J. Ubiquitin signals autophagic
degradation of cytosolic proteins and peroxisomes. (2008) PNAS . 105 (52): 20567-74.
61


32 PEX

Smith and Aitchison, ature Reviews, Molecular Cell

Biology 14:803, Dec 2013

62
 / -

Peroxisomal disorders

) (single peroxisomal enzyme deficiencies, PEDs)

) (eroxisome biogenesis disorders, PBDs).

1:5000
: X-linked adrenoleukodystrophy ()

63
 Y-

H2O2,
: (
)

1) (- )
2) . H2O2
3) -
4)
5)
6)

- ()
64