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Imaging lipids using lipid-binding toxins
Published on Sep 28, 2018852 Views
Biomembranes exhibit specific lipid organization, such as transbilayer asymmetry and domain structures. However, detailed organization of lipids is not well understood mainly because of the lack of
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Imaging lipids using lipid-binding toxins - 100:00
Imaging lipids using lipid-binding toxins - 200:21
Imaging lipids using lipid-binding toxins - 300:29
Lipids are not randomly distributed in the cell - 100:53
Lipids are not randomly distributed in the cell - 201:20
Lipids are not randomly distributed in the cell - 301:22
Lipid raft hypothesis - 101:51
Lipid raft hypothesis - 202:16
Our goal02:26
Difficulty of imaging lipids - 102:42
Difficulty of imaging lipids - 202:49
Difficulty of imaging lipids - 303:27
Difficulty of imaging lipids - 403:34
Our attempt03:49
A toxin-based probe reveals cytoplasmic exposure of Golgi sphingomyelin.04:12
Imaging sphingomyelin04:47
Lysenin04:54
Structure of Membrane Lipids05:26
MBP(Maltose-binding protein)-Lysenin specifically recognizes sphingomyelin in ELISA06:02
Lysenin induces characteristic honeycomb structure in sphingomyelin-containing membranes06:10
Lysenin induces characteristic honeycomb structure in sphingomyelin-containing membranes06:19
Recognition of sphingomyelin by lysenin mutants06:43
Detection of sphingomyelin clusters by lysenin07:12
Sphingomyelin-rich domain localizes in caveolae08:05
Sphingomyelin-rich domain does not localize in the Golgi apparatus08:45
Sphingomyelin-rich domain localizes in percentriolar endosomes - 109:08
Sphingomyelin-rich domain localizes in percentriolar endosomes - 209:14
List of memrane lipid probes used09:23
Imaging Lipid Asymmetry10:17
Asymmetric distribution of lipids in plasma membrane10:30
phosoholipase11:17
SDS-digested 13:08
Characteristics of SDS-FRL method in lipid biology - 114:50
Characteristics of SDS-FRL method in lipid biology - 215:56
Lips are not scrambled during sample preparation17:36
Sphingomyelin (SM) labeling in human erythrocyte membrane18:32
Phosphatidylcholine (PC) labeling in human erythrocyte membrane18:48
Phosphatidylethanolamine (PE) labeling in human erythrocyte membrane18:53
Phosphatidylserine (PS)/ phosphatidylinositol (PI) labeling in human erythrocyte membrane18:59
Phosphatidylinositol- 4,5-bisphosphate (PIP2) labelling in human erythrocyte membrane19:07
Revisiting asymmetrical distribution of phospholipids in the human erythrocyte membrane19:16
Distribution of phosphatidylethanolamine in the plasma membrane of fibroblast19:58
Distribution of sphingomyelin in the plasma membrane of fibroblast20:16
Distribution of sphingomyelin in the plasma membrane of neutrophils (E-face (outer leaflet))20:33
Distribution of sphingomyelin in the plasma membrane of neutrophils (P-face (inner leaflet))20:42
Distribution of phospholipids in the plasma membrane from human skin fibroblasts20:52
Transbilayer colocalization of sphingomyelin-rich domain and PIP5Kb21:15
Communication between outer and inner leaflet21:41
Outer leaflet sphingomyelin and inner leaflet sphingomyelin do not co-localize22:17
Simultaneous observation of inner leaflet and outer leaflet lipids23:29
Colocalization of sphingomyelin and PIP224:03
Photoactivation localization microscopy (PALM) image of cholesterol-rich membrane domains24:26
Sphingomyelin-rich domains colocalize with PIP2 and are required to maintain PIP2 domains25:51
Addition of exogenous sphingomyelin restores the PIP2 domain in sphingomyelinase-treated cells27:12
Transbilayer colocalization of sphingomyelin-rich domain and PIP5Kb27:30
Summary 228:08
Interbilayer co-localization of sphingomyelin and PIP2 - 128:24
Interbilayer co-localization of sphingomyelin and PIP2 - 228:39
Role of sphingomyelin in cell division28:45
Sphingomyelin is accumulated to the cleavage furrow during cytokinesis28:58
Sphingomyelinase treatment inhibits the completion of cytokinesis29:33
Sphingomyelinase treatment results in the inhibition of the completion of cytokinesis29:50
Addition of exogenous sphingomyelin restores cell division in sphingomyelinase-treated cells30:00
Sphingomyelin is accumulated to the cleavage furrow during cell divison30:08
Accumulation of phosphatidylinositol 4,5-bisphosphate to the cleavage furrow is crucial for cytokinesis30:11
Sphingomyelinase treatment abolishes the accumulation of PIP2 to the cleavage furrow30:37
Sphingomyelinase treatment abolishes the accumulation of RhoA to the cleavage furrow31:04
Summary 331:34
Role of sphingomyelin-PIP2 interaction in virus budding31:58
HIV-1 (human immunodeficiency virus type 1)32:14
Lipids incorporated in HIV-1 particles32:55
Gag protein in virus formation33:06
Gag assembly in plasma membrane33:30
EGFP-Lys and Gag-mCherry observed by confocal microscope33:52
PALM/STORM image of Gag-mEos2 and Alexa Fluor647-NT-Lys34:05
Does the size of SM cluster change in the presence of Gag cluster?34:29
SM cluster in proximity of Gag cluster is larger than that without Gag cluster 34:37
FRAP (fluorescent recovery after photobleaching)35:07
FRAP experiment of EGFP-NT-Lys35:52
Summary: FRAP experiment of EGFP-NT-Lys36:13
Summary 436:25
How does inner leaflet Gag affect outer leaflet Sphingomyelin?36:32
Sphingomyelin36:50
Sphingomyelin has unique fatty acid composition37:38
Interdigitation of lipids could link inner leaflet lipids to the outer leaflet lipids38:20
De novo synthesis patway of spingolipids38:40
PALM/STORM imaging of Gag and SM in CERS2 KO cells39:25
CERS2 KO39:45
Summary 540:11
Mitsuhiro Abe40:23