TY - JOUR
T1 - Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis
AU - Makino, Asami
AU - Abe, Mitsuhiro
AU - Murate, Motohide
AU - Inaba, Takehiko
AU - Yilmaz, Neval
AU - Hullin-Matsuda, Françoise
AU - Kishimoto, Takuma
AU - Schieber, Nicole L.
AU - Taguchi, Tomohiko
AU - Arai, Hiroyuki
AU - Anderluh, Gregor
AU - Parton, Robert G.
AU - Kobayashi, Toshihide
N1 - Publisher Copyright:
© FASEB.
PY - 2015/2/1
Y1 - 2015/2/1
N2 - Sphingomyelin (SM) is a major sphingolipid in mammalian cells and is reported to form specific lipid domains together with cholesterol. However, methods to examine the membrane distribution of SM are limited. We demonstrated in model membranes that fluorescent protein conjugates of 2 specific SM-binding toxins, lysenin (Lys) and equinatoxin II (EqtII), recognize different membrane distributions of SM; Lys exclusively binds clustered SM, whereas EqtII preferentially binds dispersed SM. Freeze-fracture immunoelectron microscopy showed that clustered but not dispersed SM formed lipid domains on the cell surface. Glycolipids and the membrane concentration of SM affect the SM distribution pattern on the plasma membrane. Using derivatives of Lys and EqtII as SM distribution-sensitive probes, we revealed the exclusive accumulation of SM clusters in the midbody at the time of cytokinesis. Interestingly, apical membranes of differentiated epithelial cells exhibited dispersed SM distribution, whereas SM was clustered in basolateral membranes. Clustered but not dispersed SM was absent from the cell surface of acid sphingomyelinase-deficient Niemann-Pick type A cells. These data suggest that both the SM content and membrane distribution are crucial for pathophysiological events bringing therapeutic perspective in the role of SM membrane distribution.
AB - Sphingomyelin (SM) is a major sphingolipid in mammalian cells and is reported to form specific lipid domains together with cholesterol. However, methods to examine the membrane distribution of SM are limited. We demonstrated in model membranes that fluorescent protein conjugates of 2 specific SM-binding toxins, lysenin (Lys) and equinatoxin II (EqtII), recognize different membrane distributions of SM; Lys exclusively binds clustered SM, whereas EqtII preferentially binds dispersed SM. Freeze-fracture immunoelectron microscopy showed that clustered but not dispersed SM formed lipid domains on the cell surface. Glycolipids and the membrane concentration of SM affect the SM distribution pattern on the plasma membrane. Using derivatives of Lys and EqtII as SM distribution-sensitive probes, we revealed the exclusive accumulation of SM clusters in the midbody at the time of cytokinesis. Interestingly, apical membranes of differentiated epithelial cells exhibited dispersed SM distribution, whereas SM was clustered in basolateral membranes. Clustered but not dispersed SM was absent from the cell surface of acid sphingomyelinase-deficient Niemann-Pick type A cells. These data suggest that both the SM content and membrane distribution are crucial for pathophysiological events bringing therapeutic perspective in the role of SM membrane distribution.
KW - Lipid binding protein
KW - Lipid raft
KW - Membrane lipids
KW - Sphingolipid
KW - Toxins
UR - http://www.scopus.com/inward/record.url?scp=84928562086&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84928562086&partnerID=8YFLogxK
U2 - 10.1096/fj.13-247585
DO - 10.1096/fj.13-247585
M3 - Article
C2 - 25389132
AN - SCOPUS:84928562086
VL - 29
SP - 477
EP - 493
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 2
ER -