Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells

被引:27
作者
Cronin, Ciaran N. [1 ]
Lim, Kheng B. [1 ]
Rogers, Joe [1 ]
机构
[1] Takeda San Diego, San Diego, CA 92121 USA
关键词
selenomethionine; baculovirus; insect cells; MAD phasing; multiwavelength anomalous diffraction;
D O I
10.1110/ps.072931407
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A protocol is described for the production of both intracellularly expressed and secreted selenome-thionyl-derivatized recombinant proteins in baculovirus-infected insect cells. The method results in the production of recombinant soluble proteins with an SeMet occupancy of similar to 75% and with a recovery of similar to 20% that of native protein expression. The method is independent of the percentage methionine content of the protein and is reliable and consistent. Similar results are obtained using either Spodoptera frugiperda Sf9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.
引用
收藏
页码:2023 / 2029
页数:7
相关论文
共 31 条
  • [1] Crystal structure of human dipeptidyl peptidase IV in complex with a decapeptide reveals details on substrate specificity and tetrahedral intermediate formation
    Aertgeerts, K
    Ye, S
    Tennant, MG
    Kraus, ML
    Rogers, J
    Sang, BC
    Skene, RJ
    Webb, DR
    Prasad, GS
    [J]. PROTEIN SCIENCE, 2004, 13 (02) : 412 - 421
  • [2] Eukaryotic expression: developments for structural proteomics
    Aricescu, A. R.
    Assenberg, R.
    Bill, R. M.
    Busso, D.
    Chang, V. T.
    Davis, S. J.
    Dubrovsky, A.
    Gustafsson, L.
    Hedfalk, K.
    Heinemann, U.
    Jones, I. M.
    Ksiazek, D.
    Lang, C.
    Maskos, K.
    Messerschmidt, A.
    Macieira, S.
    Peleg, Y.
    Perrakis, A.
    Poterszman, A.
    Schneider, G.
    Sixma, T. K.
    Sussman, J. L.
    Sutton, G.
    Tarboureich, N.
    Zeev-Ben-Mordehai, T.
    Jones, E. Yvonne
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2006, 62 : 1114 - 1124
  • [3] A time- and cost-efficient system for high-level protein production in mammalian cells
    Aricescu, A. Radu
    Lu, Weixian
    Jones, E. Yvonne
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2006, 62 : 1243 - 1250
  • [4] Highly efficient selenomethionine labeling of recombinant proteins produced in mammalian cells
    Barton, William A.
    Tzvetkova-Robev, Dorothea
    Erdjument-Bromage, Hediye
    Tempst, Paul
    Nikolov, Dimitar B.
    [J]. PROTEIN SCIENCE, 2006, 15 (08) : 2008 - 2013
  • [5] Producing selenomethionine-labeled proteins with a baculovirus expression vector system
    Bellizzi, JJ
    Widom, J
    Kemp, CW
    Clardy, J
    [J]. STRUCTURE, 1999, 7 (11) : R263 - R267
  • [6] Brodsky Oleg, 2006, Journal of Structural and Functional Genomics, V7, P101, DOI 10.1007/s10969-006-9013-0
  • [7] Selenomethionine incorporation in Saccharomyces cerevisiae RNA polymerase II
    Bushnell, DA
    Cramer, P
    Kornberg, RD
    [J]. STRUCTURE, 2001, 9 (01) : R11 - R14
  • [8] Crystallization and preliminary diffraction studies of the ectodomain of the envelope glycoprotein D from herpes simplex virus 1 alone and in complex with the ectodomain of the human receptor HveA
    Carfí, A
    Gong, HY
    Lou, H
    Willis, SH
    Cohen, GH
    Eisenberg, RJ
    Wiley, DC
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2002, 58 : 836 - 838
  • [9] CHEN WY, 1991, J BIOL CHEM, V266, P8192
  • [10] CHEN WY, 1991, J BIOL CHEM, V266, P9355