Abstract
Dynamin is a 100-kDa GTPase with multiple domains. Some of these have known functions, namely, the N-terminal GTPase domain, the PH domain that binds phosphatidylinositol lipids, and the C-terminal proline-arginine-rich domain (PRD) that binds to several SH3 domain-containing dynamin partners. Others, for example, the "middle" located between the GTPase domain and the PH domain and a predicted α-helical domain located between the PH domain and PRD, have unknown functions. Dynamin exists as a homotetramer in solution and self-assembles into higher-order structures resembling rings and helical stacks of rings. Dynamin self-assembly stimulates its GTPase activity. We used limited proteolysis to dissect dynamin's domain structure and to gain insight into intradomain interactions that regulate dynamin self-assembly and stimulate GTPase activity. We found that the PH domain functions as a negative regulator of dynamin self-assembly and stimulates GTPase activity and that the α-helical domain, termed GED for GTPase effector domain, is required for stimulated GTPase activity.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 475-483 |
| Number of pages | 9 |
| Journal | Methods |
| Volume | 20 |
| Issue number | 4 |
| DOIs | |
| State | Published - 2000 |
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ASJC Scopus subject areas
- Molecular Biology
Cite this
Domain structure and function of dynamin probed by limited proteolysis. / Muhlberg, Amy B.; Schmid, Sandra L.
In: Methods, Vol. 20, No. 4, 2000, p. 475-483.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Domain structure and function of dynamin probed by limited proteolysis
AU - Muhlberg, Amy B.
AU - Schmid, Sandra L.
PY - 2000
Y1 - 2000
N2 - Dynamin is a 100-kDa GTPase with multiple domains. Some of these have known functions, namely, the N-terminal GTPase domain, the PH domain that binds phosphatidylinositol lipids, and the C-terminal proline-arginine-rich domain (PRD) that binds to several SH3 domain-containing dynamin partners. Others, for example, the "middle" located between the GTPase domain and the PH domain and a predicted α-helical domain located between the PH domain and PRD, have unknown functions. Dynamin exists as a homotetramer in solution and self-assembles into higher-order structures resembling rings and helical stacks of rings. Dynamin self-assembly stimulates its GTPase activity. We used limited proteolysis to dissect dynamin's domain structure and to gain insight into intradomain interactions that regulate dynamin self-assembly and stimulate GTPase activity. We found that the PH domain functions as a negative regulator of dynamin self-assembly and stimulates GTPase activity and that the α-helical domain, termed GED for GTPase effector domain, is required for stimulated GTPase activity.
AB - Dynamin is a 100-kDa GTPase with multiple domains. Some of these have known functions, namely, the N-terminal GTPase domain, the PH domain that binds phosphatidylinositol lipids, and the C-terminal proline-arginine-rich domain (PRD) that binds to several SH3 domain-containing dynamin partners. Others, for example, the "middle" located between the GTPase domain and the PH domain and a predicted α-helical domain located between the PH domain and PRD, have unknown functions. Dynamin exists as a homotetramer in solution and self-assembles into higher-order structures resembling rings and helical stacks of rings. Dynamin self-assembly stimulates its GTPase activity. We used limited proteolysis to dissect dynamin's domain structure and to gain insight into intradomain interactions that regulate dynamin self-assembly and stimulate GTPase activity. We found that the PH domain functions as a negative regulator of dynamin self-assembly and stimulates GTPase activity and that the α-helical domain, termed GED for GTPase effector domain, is required for stimulated GTPase activity.
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U2 - 10.1006/meth.2000.0960
DO - 10.1006/meth.2000.0960
M3 - Article
VL - 20
SP - 475
EP - 483
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
IS - 4
ER -