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本帖最后由 细胞海洋 于 2013-5-6 09:12 编辑 f0 J1 P; T* L2 P
% T+ V& {5 R- i3 ONanoCellBiology of Secretion
' D: @1 i' I6 BImaging Its Cellular and Molecular Underpinnings
2 Q/ B# j0 k5 Q# I1 b5 e$ r
" Z: i; h( ?# CNanoCellBiology of Secretion ....................................................................... 1
. w' i2 J& q/ cIntroduction ...................................................................................................... 1% H2 V6 ~5 l. u5 t" h
Materials and Methods ..................................................................................... 51 O; r/ K7 z# y8 K; X
Pancreatic Acini Isolation ............................................................................ 5* B. J' F. O: \7 K7 u
Zymogen Granule Isolation ......................................................................... 5( _6 @1 w6 w" q! M7 L4 F
Isolation of Synaptosomes, Synaptosomal Membrane,
, t6 U5 a- n4 Fand Synaptic Vesicles ................................................................................... 6/ Q! K, E, X% k: k* u
Porosome Isolation ....................................................................................... 6" O& f: i9 M6 \& p0 v) n
Atomic Force Microscopy ........................................................................... 7
6 U, E* F+ ` M8 W3 T& wMeasurement of Secretion ........................................................................... 70 \6 J D1 ?- x! l" A/ `. h+ j) @
Transmission Electron Microscopy.............................................................. 8( \# V1 `' F1 J9 V
Dynamic Light Scattering (DLS) to Determine Vesicle' Q8 d3 L9 u! w2 L
Volume Change ............................................................................................ 80 w* F) N6 x0 A9 M; J, }
Photon Correlation Spectroscopy ................................................................ 8; `! D7 ? o- B, c) i
Porosome Functional Reconstitution Assay ................................................ 9
. Y6 y+ m; s7 @0 ]Preparation of Liposomes and SNARE Reconstitutions .............................. 9
5 Q1 C9 G: A7 l! OCircular Dichroism Spectroscopy ................................................................ 103 g9 U' v2 C0 S, E
Wide-Angle X-ray Diffraction ..................................................................... 10
. m0 i. x0 D! |( R/ o TPorosome Discovery ........................................................................................ 119 J4 }% \$ K0 g% x2 `% a! ?
Calcium and SNARE-Induced Membrane Fusion ........................................... 23
* n' L$ F# N* }# d: x1 k5 Z: Wv- and t-SNAREs Need to Reside in Opposing Membrane0 j7 r2 n) x( z( u, C# A: X9 o4 w
for Appropriate Interactions and the Establishment
3 o8 `" Z3 x$ u, nof Continuity Between Opposing Membranes ............................................. 265 a: W9 e2 [: i* R
Membrane Curvature Dictates the Size of the SNARE- w. ]8 d+ E2 N
Ring Complex .............................................................................................. 31 P8 L/ t ~; N" n* k, S4 T
Disassembly of the t-/v-SNARE Complex .................................................. 32- d/ R0 n! Z7 U8 D
CD Spectroscopy Confi rms Membrane Requirement* ?& t+ ^2 W/ i7 j! i. K: G
for Appropriate t-/v-SNARE Assembly and Suffi ciency
9 v' F% o9 {1 g3 Z+ nof NSF–ATP for Its Disassembly ................................................................. 38 I D, J% E' v" r( F9 y$ C
SNAREs Bring Opposing Bilayers into Close Approximation,
; I' y, b. J' R& p7 @5 uEnabling Calcium Bridging and Membrane Fusion .................................... 40
2 M* q$ j: X5 l& p/ z8 m8 O9 h6 }+ C" ]Membrane Lipids Infl uence SNARE Complex0 T) K$ Q4 I& [$ F& f: q
and Assembly–Disassembly ........................................................................ 42
* D. P7 [. ~, q- FSecretion Involves Vesicle Swelling and Content Expulsion........................... 47
: e0 _0 \. n3 \ mSecretory Vesicle Swelling Is Required for the Expulsion" y8 f/ q1 Z8 d* I2 F# f
of Intravesicular Contents During Cell Secretion ........................................ 47
& B( b. Y1 ^* P3 ^4 I2 {6 y6 BMolecular Mechanism of Secretory Vesicle Swelling ................................. 53
( Q$ W3 H+ p5 f9 w, ~8 _+ dPresence of Functional b-Adrenergic Receptors at the Synaptic
; I. d) T- Z3 q$ J0 U5 S7 |4 IVesicle Membrane ........................................................................................ 58! g& b' I4 H0 s. O5 ^) y
Conclusion ....................................................................................................... 62
0 I. B+ C; I1 u5 v2 OReferences ........................................................................................................ 63
9 H6 v0 a/ ]6 A3 VIndex ................................................................................................................ 69
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发表于 2013-5-5 21:52
