Crystallization of membrane proteins in Cubo

P. Nollert, Javier Navarro, E. M. Landau

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Our understanding of lipidic cubic phases for the crystallization of membrane proteins has advanced greatly since the inception of the concept in 1996, and the method is becoming well accepted . Several protocols that allow the efficient screening of crystallization conditions and handling of crystals are presented. State-of-the-art micro techniques allow a large number of crystallization conditions to be tested using very small amounts of protein, and diffraction quality crystals can be grown in larger volumes in glass vials. In cubo crystallization conditions differ from those employed for detergent-solubilized proteins. Variations comprise the type of lipid matrix, detergent, protein, salt, temperature, hydration, pH, and pressure. Commercially available screening kits may be applied in order to define lead conditions. Once obtained, crystals may be removed from the surrounding cubic phase mechanically, by enzymatic hydrolysis, or by detergent solubilization. We anticipate this set of protocols to be applied successfully to larger, less stable, and noncolored membrane proteins in order to obtain well-diffracting crystals of membrane proteins that have so far evaded crystallization in the detergent-solubilized state.

Original languageEnglish
Pages (from-to)183-199
Number of pages17
JournalMethods in Enzymology
Volume343
DOIs
StatePublished - 2001
Externally publishedYes

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Crystallization
Membrane Proteins
Detergents
Crystals
Screening
Proteins
Enzymatic hydrolysis
Hydration
Glass
Hydrolysis
Salts
Diffraction
Lipids
Pressure
Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystallization of membrane proteins in Cubo. / Nollert, P.; Navarro, Javier; Landau, E. M.

In: Methods in Enzymology, Vol. 343, 2001, p. 183-199.

Research output: Contribution to journalArticle

Nollert, P. ; Navarro, Javier ; Landau, E. M. / Crystallization of membrane proteins in Cubo. In: Methods in Enzymology. 2001 ; Vol. 343. pp. 183-199.
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