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本帖最后由 细胞海洋 于 2013-5-6 09:12 编辑
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& i3 m+ V4 a3 C. l) b2 T( b+ Z! e+ CNanoCellBiology of Secretion. d. _7 p5 h; |/ \% C* J# Y" {
Imaging Its Cellular and Molecular Underpinnings
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2 r. o1 Y6 c0 k! L ANanoCellBiology of Secretion ....................................................................... 17 X- w- I h+ Z% v. u
Introduction ...................................................................................................... 1
$ D+ [9 K* |$ L+ xMaterials and Methods ..................................................................................... 5
9 ^+ w, [3 \' mPancreatic Acini Isolation ............................................................................ 5
- S! {+ g, t6 |$ w: `Zymogen Granule Isolation ......................................................................... 5! ?1 h) I& [- A: Q6 b
Isolation of Synaptosomes, Synaptosomal Membrane,
4 X4 e7 o6 X5 g: {0 Zand Synaptic Vesicles ................................................................................... 6
% |$ c9 }& h6 j) yPorosome Isolation ....................................................................................... 6
4 C+ B. }! c" a6 I* c8 q4 cAtomic Force Microscopy ........................................................................... 7" T0 v3 T7 f" k9 C
Measurement of Secretion ........................................................................... 7
+ V- D, Z4 v: F% G1 ] V- bTransmission Electron Microscopy.............................................................. 8
. ?# ?5 s+ A# S; D% l! HDynamic Light Scattering (DLS) to Determine Vesicle
/ v6 G0 l7 {* P& j& eVolume Change ............................................................................................ 8
( m/ v- A2 l7 i# j% ^& ^7 G; ZPhoton Correlation Spectroscopy ................................................................ 8
; \2 S: f, S/ |& J, h9 K+ BPorosome Functional Reconstitution Assay ................................................ 9
5 G p0 J! @7 _0 Y* |1 @Preparation of Liposomes and SNARE Reconstitutions .............................. 9
" R+ b/ v9 P/ w9 y* I$ z2 ]Circular Dichroism Spectroscopy ................................................................ 10
`" a) t% W6 Y/ t' I1 C, dWide-Angle X-ray Diffraction ..................................................................... 10
: w" B% E8 E5 b/ G6 YPorosome Discovery ........................................................................................ 11
6 {; X9 X1 O$ B* M* D+ \ ^8 l$ tCalcium and SNARE-Induced Membrane Fusion ........................................... 23
+ U( H! p. x/ Tv- and t-SNAREs Need to Reside in Opposing Membrane
# E+ ^7 w5 n: w h. q# Ifor Appropriate Interactions and the Establishment: y- p; M# t# l# N
of Continuity Between Opposing Membranes ............................................. 263 E" I- V' @: _( j. ~7 k+ @9 F6 v! f
Membrane Curvature Dictates the Size of the SNARE
7 L' `" d1 D6 U9 N$ bRing Complex .............................................................................................. 31
% @4 k9 f1 G _, _% G( `4 Z: JDisassembly of the t-/v-SNARE Complex .................................................. 32
! G+ S5 r$ P$ q, jCD Spectroscopy Confi rms Membrane Requirement
C+ I7 N% s+ Gfor Appropriate t-/v-SNARE Assembly and Suffi ciency
+ Q2 G4 b. [1 s/ P9 R I3 bof NSF–ATP for Its Disassembly ................................................................. 38( e9 V9 p4 C+ U
SNAREs Bring Opposing Bilayers into Close Approximation,
; P# T8 Y3 m* x- VEnabling Calcium Bridging and Membrane Fusion .................................... 40
( H% o, x [) q6 E1 H0 OMembrane Lipids Infl uence SNARE Complex
8 T% E# X' v- Qand Assembly–Disassembly ........................................................................ 42& G( }8 t0 V& A7 a" ~
Secretion Involves Vesicle Swelling and Content Expulsion........................... 47
\2 l7 H" I- B+ N; A2 Y6 u8 LSecretory Vesicle Swelling Is Required for the Expulsion# X1 c- G, Y/ j' c2 w! a! o6 T
of Intravesicular Contents During Cell Secretion ........................................ 473 f8 S3 G' ?0 l" Q6 p2 M
Molecular Mechanism of Secretory Vesicle Swelling ................................. 53, H, l* K& {. a+ Q+ r
Presence of Functional b-Adrenergic Receptors at the Synaptic
- w' ?: K+ f! w' l" U" Z9 Y$ `/ EVesicle Membrane ........................................................................................ 588 i% p- B0 K1 J; U
Conclusion ....................................................................................................... 62
1 @& }3 |0 u' M0 w; E2 DReferences ........................................................................................................ 634 U/ T+ ?8 t. Z
Index ................................................................................................................ 693 o8 s* |$ M) l$ H
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发表于 2013-5-5 21:52
