Dynamin regulates focal exocytosis in phagocytosing macrophages

被引:55
作者
Di, AK [1 ]
Nelson, DJ [1 ]
Bindokas, V [1 ]
Brown, ME [1 ]
Libunao, F [1 ]
Palfrey, HC [1 ]
机构
[1] Univ Chicago, Dept Neurobiol Pharmacol & Physiol, Chicago, IL 60637 USA
关键词
D O I
10.1091/mbc.E02-09-0626
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Phagocytosis in macrophages is thought to involve insertion of cytoplasmic vesicles at sites of membrane expansion before particle ingestion ("focal" exocytosis). Capacitance (Cm) measurements of cell surface area were biphasic, with an initial rise indicative of exocytosis followed by a fall upon phagoctosis. Unlike other types of regulated exocytosis, the Cm rise was insensitive to intracellular Ca2+, but was inhibited by guanosine 5'-O-(2-thio)diphosphate. Particle uptake, but not Cm rise, was affected by phosphatidylinositol 3-kinase inhibitors. Inhibition of actin polymerization eliminated the Cm rise, suggesting possible coordination between actin polymerization and focal exocytosis. Introduction of anti-pan-dynamin IgG blocked Cm changes, suggesting that dynamin controls focal exocytosis and thereby phagocytosis. Similarly, recombinant glutathione S-transferase.amphiphysin-SH3 domain, but not a mutated form that cannot bind to dynamin, inhibited both focal exocytosis and phagocytosis. Immunochemical analysis of endogenous dynamin distribution in macrophages revealed a substantial particulate pool, some of which localized to a presumptive endosomal compartment. Expression of enhanced green fluorescent protein.dynamin-2 showed a motile dynamin pool, a fraction of which migrated toward and within the phagosomal cup. These results suggest that dynamin is involved in the production and/or movement of vesicles from an intracellular organelle to the cell surface to support membrane expansion around the engulfed particle.
引用
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页码:2016 / 2028
页数:13
相关论文
共 49 条
[21]   The SH3 domain of amphiphysin binds the proline-rich domain of dynamin at a single site that defines a new SH3 binding consensus sequence [J].
Grabs, D ;
Slepnev, VI ;
Zhou, SY ;
David, C ;
Lynch, M ;
Cantley, LC ;
DeCamilli, P .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (20) :13419-13425
[22]  
Greenberg S, 2001, J CELL SCI, V114, P1039
[23]  
GREENBERG S, 1999, INFLAMMATION BASIC P, P681
[24]   v-SNARE-dependent secretion is required for phagocytosis [J].
Hackam, DJ ;
Rotstein, OD ;
Sjolin, C ;
Schreiber, AD ;
Trimble, WS ;
Grinstein, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (20) :11691-11696
[25]   Membrane capacitance changes associated with particle uptake during phagocytosis in macrophages [J].
Holevinsky, KO ;
Nelson, DJ .
BIOPHYSICAL JOURNAL, 1998, 75 (05) :2577-2586
[26]   DIFFERENCES IN MODE OF PHAGOCYTOSIS WITH FC AND C3 RECEPTORS IN MACROPHAGES [J].
KAPLAN, G .
SCANDINAVIAN JOURNAL OF IMMUNOLOGY, 1977, 6 (08) :797-807
[27]  
Kwiatkowska K, 1999, BIOESSAYS, V21, P422, DOI 10.1002/(SICI)1521-1878(199905)21:5<422::AID-BIES9>3.0.CO
[28]  
2-#
[29]   Functional separation of pseudopod extension and particle internalization during Fcγ receptor-mediated phagocytosis [J].
Lowry, MB ;
Duchemin, AM ;
Robinson, JM ;
Anderson, CL .
JOURNAL OF EXPERIMENTAL MEDICINE, 1998, 187 (02) :161-176
[30]  
May RC, 2001, J CELL SCI, V114, P1061