A Practical Method for Uniform Isotopic Labeling of Recombinant Proteins in Mammalian Cells

Andrew P. Hansen, Andrew M. Petros, Andrew P. Mazar, Terry M. Pederson, Annemarie Rueter, Stephen W. Fesik

Research output: Contribution to journalArticlepeer-review

65 Scopus citations

Abstract

A method to obtain uniformly isotopically labeled (15N and 15N/13C) protein from mammalian cells is described. The method involves preparation of isotopically labeled media consisting of amino acids isolated from bacterial and algal extracts supplemented with cysteine and enzymatically synthesized glutamine. The approach is demonstrated by producing 15N-labeled and 15N/13C-labeled urokinase from Sp2/0 cells and successfully growing Chinese hamster ovary (CHO) cells on the labeled media. Thus, using the procedures described, isotopically labeled proteins that have been expressed in mammalian cells can be prepared, allowing them to be studied by heteronuclear multidimensional NMR techniques.

Original languageEnglish (US)
Pages (from-to)12713-12718
Number of pages6
JournalBiochemistry
Volume31
Issue number51
DOIs
StatePublished - Feb 1 1992

ASJC Scopus subject areas

  • Biochemistry

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