Protein engineering for crystallization of the GTPase Sar1 that regulates ER vesicle budding.

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Bibliographic Details
Title:	Protein engineering for crystallization of the GTPase Sar1 that regulates ER vesicle budding.
Authors:	Mingdong Huang^1,2,3, Weissman, Jacques T.¹, Chenqian Wang¹, Balch, William E.^1,2,4 webalch@scripps.edu, Wilson, Lan A.^2,5
Source:	Acta Crystallographica: Section D (Wiley-Blackwell). Apr2002, Vol. 58 Issue 4, p700-703. 4p.
Subjects:	Protein engineering, Crystallization, Light scattering, Biochemical engineering, Genetic engineering, Guanosine triphosphatase
Abstract:	Sar1 is an important and unique GTPase that regulates vesicle budding from the ER membrane. An effort to crystallize full-length hamster Sar1 was unsuccessful owing to the aggregation of Sar1 in solution as indicated by dynamic light-scattering measurements. It was presumed that a patch of hydrophobic residues in the N-terminal region of Sar1 was responsible for the aggregation. To attempt to improve protein crystallizability, the N-terminal residues of Sari were progressively truncated and the solution behavior of the resulting Sari variants was monitored by dynamic light scattering. Truncation of the first nine residues from the N-terminus led to a Sar1 variant that is monodisperse in solution. This well behaved Sari variant yielded crystals in just a few days that were ultimately refined to diffraction quality. [ABSTRACT FROM AUTHOR]
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Database:	Engineering Source

Description
Abstract:	Sar1 is an important and unique GTPase that regulates vesicle budding from the ER membrane. An effort to crystallize full-length hamster Sar1 was unsuccessful owing to the aggregation of Sar1 in solution as indicated by dynamic light-scattering measurements. It was presumed that a patch of hydrophobic residues in the N-terminal region of Sar1 was responsible for the aggregation. To attempt to improve protein crystallizability, the N-terminal residues of Sari were progressively truncated and the solution behavior of the resulting Sari variants was monitored by dynamic light scattering. Truncation of the first nine residues from the N-terminus led to a Sar1 variant that is monodisperse in solution. This well behaved Sari variant yielded crystals in just a few days that were ultimately refined to diffraction quality. [ABSTRACT FROM AUTHOR]
ISSN:	09074449
DOI:	10.1107/S090744490200238X