Crystallization of the yeast MATα2/MCM1/DNA ternary complex: General methods and principles for protein/DNA cocrystallization

Song Tan, Yvonne Hunziker, Luca Pellegrini, Timothy J. Richmond

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

We describe our efforts to crystallize binary MCM1/DNA and ternary MATα2/MCM1/DNA complexes, including the unsuccessful attempts to crystallize MCM1/DNA complexes and the successful design of DNA crystal packing that resulted in high-resolution crystals of the MATα2/MCM1/DNA complex. We detail general procedures useful for preparing protein/DNA cocrystals, including improved methods for producing and purifying DNA-binding proteins and DNA fragments, for purifying protein/DNA complexes, and for controlling pH conditions during crystallization. We also describe the rational design of DNA for protein/DNA cocrystallization attempts, based on our analysis of how straight and bent DNA with single base-pair overhangs can pack end-to-end in a crystal. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)947-959
Number of pages13
JournalJournal of Molecular Biology
Volume297
Issue number4
DOIs
StatePublished - Apr 7 2000

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

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