PDF电子书：NanoCellBiology of Secretion

zsg

- ]. o7 t8 q" ]  d  b1 L+ K9 t$ V
+ c5 b2 [, }& D4 f8 {& p
NanoCellBiology of Secretion
9 \/ G/ D% @  OImaging Its Cellular and Molecular Underpinnings  [4 Y1 o7 @4 u* h+ \& c
- `; q+ e0 [+ Z* I" C
NanoCellBiology of Secretion ....................................................................... 1' p% S3 z* S4 B: ]4 [0 k5 D+ b7 V) u. j+ m
Introduction ...................................................................................................... 1+ ~" C/ i6 ^! P  Q, \
Materials and Methods ..................................................................................... 5
- ?& O7 r+ g; Q6 T) vPancreatic Acini Isolation ............................................................................ 5
, L7 g: P+ o0 }; O, s- j) @Zymogen Granule Isolation ......................................................................... 57 a: f% A- N% ?: m
Isolation of Synaptosomes, Synaptosomal Membrane,7 w+ H: g3 U2 E2 H4 E3 p/ |& ?
and Synaptic Vesicles ................................................................................... 6; ~6 o; j( o$ {# k
Porosome Isolation ....................................................................................... 6* w$ n, V" t$ E7 \& X
Atomic Force Microscopy ........................................................................... 7/ w3 L; o; Z( R4 Y1 c
Measurement of Secretion ........................................................................... 7
+ @" N% M4 c5 |, v3 h2 DTransmission Electron Microscopy.............................................................. 8# _" H/ \' X7 d# {; u" L
Dynamic Light Scattering (DLS) to Determine Vesicle
- C+ \$ y' p  C& Z' XVolume Change ............................................................................................ 8/ o0 I& d/ y# e. G6 A5 a- v
Photon Correlation Spectroscopy ................................................................ 8
8 J" v* n# O; T7 wPorosome Functional Reconstitution Assay ................................................ 9
5 d% L3 ]4 L2 O, ?2 m. ]Preparation of Liposomes and SNARE Reconstitutions .............................. 9# M" t$ k. r" H. @
Circular Dichroism Spectroscopy ................................................................ 102 g) g& e* }7 c& N
Wide-Angle X-ray Diffraction ..................................................................... 10) \# ~* P, {) P2 k9 J% l6 E
Porosome Discovery ........................................................................................ 11# s) B6 k. O1 F
Calcium and SNARE-Induced Membrane Fusion ........................................... 233 S5 V; ?2 F. K: T2 j7 }
v- and t-SNAREs Need to Reside in Opposing Membrane
3 Y5 _2 W9 _2 \for Appropriate Interactions and the Establishment
- q$ M& Z$ f1 U' u  a# kof Continuity Between Opposing Membranes ............................................. 262 v: g2 O' e3 h2 k9 o! f6 ~
Membrane Curvature Dictates the Size of the SNARE6 y  ]' h6 \/ _6 m
Ring Complex .............................................................................................. 31
: \5 n- W! W% d; ~" S. J- s3 IDisassembly of the t-/v-SNARE Complex .................................................. 328 z3 |  h" N. W
CD Spectroscopy Confi rms Membrane Requirement2 T' t3 G4 E9 S' d* v6 Z0 t
for Appropriate t-/v-SNARE Assembly and Suffi ciency
) V0 S  K0 f3 k) Aof NSF–ATP for Its Disassembly ................................................................. 382 q, I( X: ^, P4 n7 q
SNAREs Bring Opposing Bilayers into Close Approximation,
; S0 d4 [, X0 Z- CEnabling Calcium Bridging and Membrane Fusion .................................... 40
$ d2 k9 ^% V( a- H( \' i; w/ y4 CMembrane Lipids Infl uence SNARE Complex: Y" b: O8 @" C4 A) p2 g
and Assembly–Disassembly ........................................................................ 42* P5 G9 w; b- D/ J
Secretion Involves Vesicle Swelling and Content Expulsion........................... 47
8 @5 N0 a2 i6 w4 g% a5 z* b. aSecretory Vesicle Swelling Is Required for the Expulsion
; K. r, H. v" M$ l' u9 D/ _7 u- [of Intravesicular Contents During Cell Secretion ........................................ 47
4 R& b1 x+ u7 B  R! \: p8 EMolecular Mechanism of Secretory Vesicle Swelling ................................. 53
" n, H; S( {5 O$ I/ p) k1 l' JPresence of Functional b-Adrenergic Receptors at the Synaptic6 \( R$ M4 L. e. t1 }
Vesicle Membrane ........................................................................................ 58
) F4 G/ k. f* v& eConclusion ....................................................................................................... 62
# F3 Q1 C' B  O' E. K, tReferences ........................................................................................................ 63
" ~) K" m& A- i% zIndex ................................................................................................................ 694 n1 n# X9 o) ~& m
[hide][/hide]