Cilia: A
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Amsterdam [u.a.]
Elsevier
2013
|
| Ausgabe: | 1. ed. |
| Schriftenreihe: | Methods in enzymology
524 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | LIV, 397 S. Ill., graph. Darst. |
| ISBN: | 9780123979452 0123979455 |
Internformat
MARC
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| 245 | 1 | 0 | |a Cilia |n A |c ed. by Wallace F. Marshall |
| 250 | |a 1. ed. | ||
| 264 | 1 | |a Amsterdam [u.a.] |b Elsevier |c 2013 | |
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Datensatz im Suchindex
| _version_ | 1804150206286004224 |
|---|---|
| adam_text | CONTENTS
Contributors
xi
Preface
xv
Volume in Series
xix
1.
Isolation of Intraflagellar Transport Particle Proteins from
Chlamydomonas reinhardtii
1
Elizabeth Richey and Hongmin Qin
1.
Introduction
2
2.
Media
4
3.
Isolating
Flagella
5
4.
Preparing Soluble
Flagellar
Proteins
9
5.
Isolating IFT Particle Proteins by Fractioning Soluble
Flagellar
Proteins
Through Sucrose Density Gradient
9
6.
Isolating IFT Particle Proteins by Immunoprecipitatlon from Soluble
Flagellar
Proteins
14
7.
Conclusions
15
Acknowledgments
15
References
16
2.
Molecular Tools for Studying the Radial Spoke
19
Xiaoyan Zhu, Yi Liu, Priyanka Sivadas, Anjali Gupta, and Pinfen Yang
1.
Introduction
20
2.
Recovery of Genomic
DNA 21
3.
Slot Lysis Assay for Large-Scale Screenings
24
4. DNA
Alterations
26
5.
Tagging
28
6.
Ni-NTA Affinity Purification of the Radial Spoke Complex
30
7.
Flagella
Miniprep
33
8.
Summary
. 34
Acknowledgments
34
References
34
3.
Methods for Analysis of Calcium/Calmodulin Signaling
in Cilia and
Flagella
37
Christen G. DiPetrillo and Elizabeth F. Smith
1.
Introduction
38
•ji
Contents
2.
Calcium Fntry
into the Intraflagellar Space
38
3.
Calcium
Binding Proteins
39
4.
Calcium Induced Modulation of Motility
46
5.
Summary
52
References
53
4.
Analysis of
Flagellar
Protein Ubiquitination
59
Huan Long and Kaiyao Huang
1.
Introduction
60
2.
Summary of Putative Ubiquitin, Ubiquitm-Like Proteins, t [?, and E3
in Magella/Cilia
61
3.
Determination of the
Flagellar
Localization of Candidate Proteins with
Fluorescence Proteins
66
4.
Analysis of the Free Ubiquitin in
Flagella
66
5.
In Vitro LJbiquitination System of
Flagella
6.
6.
Analysis of
Flagellar
Ubiquitination During
Flagellar
Shortening
68
7.
Analysis of Hageilar Ubiquitination During Mating
/1
8.
Summary
71
Acknowledgment
72
References
72
5.
Analysis of Ciliary Import
75
John F. Dishinger, H. Lynn Kee, and
Kristen
J.
Verhey
1.
Introduction
76
2.
General
Considérations
76
3.
Analysis of the Permeability Barrier to Ciliary Entry Using Micromjection
78
4.
Analysis of Ciliary Import by Fluorescence Recovery After Photobieaching
82
Acknowledgments
87
References
87
6.
Analysis of Signaling Pathways Controlling
Flagellar
Movements in Mammalian Spermatozoa
91
Fang Xie, Edward M. Eddy, and Marco
Conti
1.
Introduction
92
2.
Spermatozoa Preparations
93
3.
Expression of sAC in a Heterologous System
95
4.
Measurement of Soluble Adenylyl Cyclase Activity
96
5.
Measurement of cAMP Accumulation in Spermatozoa
99
6.
Isolation of Macromolecuiar Complexes Including sAC
100
7.
Summary
102
Contents
vii
Acknowledgment
102
References
102
7.
Analysis of Microtubule Plus-End-Tracking Proteins in Cilia
105
Jesper
Larsen, llya Grigoriev, Anna Akhmanova, and
Lotte B. Pedersen
1.
Introduction
106
2.
Detection of EB P oteins in Cilia by fluorescence Microscopy
10/
3.
Functional Analysis of EB Proteins in Cultures of Ciliated Mammalian Cells
115
4.
Identification of tB-Binding Partners in Cilia
117
5.
Summary
, 19
Acknowledgments
120
References
120
8.
Biochemical and Physiological Analysis of Axonemal Dyneins
123
Stephen M. King
1.
Introduction
124
2.
Axonema
Dynem
Subí
omolexes from Cytoplasrr
25
3.
loca.izaiion of Axonemal
Dynem
Components in Chlamydomonos I lagella
130
4.
Ana ysis of
Dynem
Driven Motility in C.hlamydomonai
138
5.
Conclusions
143
Acknowledgment
¡43
References
143
9.
Measuring the Regulation of Dynein Activity During
Flagellar
Motility
147
Chikako Shingyoji
1.
introduction
148
2.
Materials and Methods
149
3.
Important Parameters in Beating
Flagella
15?
4.
Microtubule Sliding Induced by
Dynem
157
5.
Dynein Activation by Mechanical Force of Bending
163
6.
Outlook
166
Acknowledgments
166
References
167
10.
Analysis of Interactions Between Intraflagellar
Transport Proteins
171
Robert
H. Behal
and Douglas G. Cole
1.
Introduction
177
2.
Isolation of I FT Complexes from Isolated
Flagella
174
Contents
3.
Isolation of II
Τ
Complex
A from Whole Cell Extracts
177
4.
Architectural Analysis of
ΙΠ
Complex
В
181
5.
Bactenally Based Interaction Analysis of II
ľ
B Proteins
185
6.
Yeast-Based Interaction Analysis of
HT A
Proteins
188
7.
.Summary
190
Acknowledgments
191
References
191
11.
Analysis of Ciliary Membrane Protein Dynamics Using
SNAP Technology
195
John A. Follit and Gregory J.
Pazour
1.
Introduction
195
2.
Generation of SNAP-lagged Ciliary Targeted Proteins
197
3.
P jlse-Chase Analysis of the TrafficKinq of hbrocystri SNAP to the Cilium
198
4.
Example Protocol
201
5.
future Prospects
203
Acknowledgment
203
References
203
12.
Engineering Oscillating Microtubule Bundles
205
Timothy Sanchez and
Zvonimir
Dogic
1.
Introduction
206
2.
Purification and Preparation of Proteins
208
3.
Depletion-Driven Bundling of MT
s
212
4.
Glass Surface Treatment, Acrylamide Coating, and Flow Cell Construction
214
5.
Active MT Bundle Samples
216
6.
Protocol Qualifications and Outlook
222
Acknowledgments
223
References
223
13.
Purification of Exosome-Like Vesicles from Urine
225
Christopher Y. Chen, Marie C. Hogan, and Christopher J. Ward
1.
Introduction
226
2.
Materials
229
3.
Collecting Crude ELVs
230
4.
Removing the Tamm-
-Horsfall
Protein
233
5.
Fractionation
235
6.
Downstream Processing
238
7.
Conclusions
238
References
239
14.
Electron
Microscopy of
Flagella,
Primary Cilia, and Intraflagellar
Transport in Flat Embedded Cells
243
Michaela Rogowski, Dirk Scholz, and Stefan Geimer
1.
Introduction
244
2.
Preparing Chlamydomonai reinhardtn and Monolayers of Mammalian
Culture Ceils for Flat Embedding
24/
3.
Dehydration, Infiltration, and
(
¡at Embedding
249
4.
Preparation of Blocks and Ultramicrotomy
252
5.
Summary
254
Acknowledgments
260
References
261
15./Л
vivo Imaging of IFT in Chlamydomonas
Flagella
265
Karl F. Lechtreck
1.
Introduction
266
2.
Samp.
e Prepa ation
268
3.
Visualizing IFT by DIG
271
4.
Visualizing IFT by T|RFM
272
5.
Analysis of IFT Using Kymograms
278
6.
Open Questions and I uture Directions
280
7.
Useful Links
281
8.
Summary
282
Acknowledgements
282
References
282
16.
Measuring Rates of Intraflagellar Transport Along
Caenorhabditis
elegans
Sensory Cilia Using Fluorescence
Microscopy
285
Ingrid
Brust-Mascher,
Guangshuo
Ou,
and Jonathan M. Scholey
1.
Introduction
286
2.
Preparation of Living Transgenic
C elegans
Expressing Fluorescent
FT Proteins
287
3.
Direct Tracking of 111 Proteins Using Spinning Disc Confocal Microscopy
and kymography
292
4.
indirect Estimation of the Rates of IFT using Flourescence Recovery
After Photobleachmg and Mathematical Modeling
295
5.
Summary
300
Acknowledgments
302
References
302
17. 3D
Structural
Analysis of Flagella/Cilia by Cryo-Electron
Tomography
305
Khanh Huy Bui and Takashi Ishikawa
1.
Introduction
306
2.
Purification of f-lagella
30/
3.
Preparation of Specimens for Cryo-FT
307
4.
Cryo-Electron Microscopy and Data Acquisition
308
5.
Tomogram Reconstruction
309
6.
Subtomogram Averaging 31C
7.
Classification
3Ì8
8.
Studying the Interaction of the
Flagellar
Component
319
9.
Outlook
321
References
322
18.
Electron Tomography of IFT Particles
325
Gaia Pigino,
Francesca
Cántele,
Elisa Vannuccini, Salvatore Lanzavecchia,
Eugenio Paccagnini,
and
Pietro Lupetti
1.
Introduction
326
2.
Overview of Electron Tomography
328
3.
Sample Preparation
329
4.
Tomography Data Collection
331
5. Tomographie
Reconstruction
332
Acknowledgments
340
References
340
19.
Reconstitution
of
Flagellar
Sliding
341
Joshua
Alper,
Veikko
Geyer,
Vikram
Mukundan, and Jonathon Howard
1.
Introduction
344
2.
Cultunng Chiamydomonas Cells
344
3.
Purifications
350
4.
Reconstitution
Assays
357
5.
Summary
368
References
368
Author Index
371
Subject Index
389
|
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| id | DE-604.BV040914009 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T00:35:14Z |
| institution | BVB |
| isbn | 9780123979452 0123979455 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025893284 |
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| record_format | marc |
| series | Methods in enzymology |
| series2 | Methods in enzymology |
| spelling | Cilia A ed. by Wallace F. Marshall 1. ed. Amsterdam [u.a.] Elsevier 2013 LIV, 397 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in enzymology 524 Methods in enzymology ... Proteindesign (DE-588)4346801-9 gnd rswk-swf Zilie (DE-588)4190826-0 gnd rswk-swf Zilie (DE-588)4190826-0 s Proteindesign (DE-588)4346801-9 s DE-604 Marshall, Wallace F. Sonstige oth (DE-604)BV040914191 1 Methods in enzymology 524 (DE-604)BV000000938 524 Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025893284&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Cilia Methods in enzymology Proteindesign (DE-588)4346801-9 gnd Zilie (DE-588)4190826-0 gnd |
| subject_GND | (DE-588)4346801-9 (DE-588)4190826-0 |
| title | Cilia |
| title_auth | Cilia |
| title_exact_search | Cilia |
| title_full | Cilia A ed. by Wallace F. Marshall |
| title_fullStr | Cilia A ed. by Wallace F. Marshall |
| title_full_unstemmed | Cilia A ed. by Wallace F. Marshall |
| title_short | Cilia |
| title_sort | cilia |
| topic | Proteindesign (DE-588)4346801-9 gnd Zilie (DE-588)4190826-0 gnd |
| topic_facet | Proteindesign Zilie |
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