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Identification of UDP-linked murein precursors as contaminants in recombinant proteins of low molecular weight.

Abstract：

:The A(280)/A(260) ratio of a purified protein is frequently used as an indication of the purity of the preparation with respect to nucleic acids. We show here that for low-molecular-weight recombinant proteins purified from Escherichia coli, a low A(280)/A(260) ratio can also result from contamination with UDP-linked murein precursors derived from bacterial cell wall metabolism. Although these precursors are small molecules of molecular weight 1000-1200, they comigrate in gel filtration with recombinant human FKBP (MW 11,820). This gel filtration behavior, which is distinct from that of unmodified mononucleotides, does not reflect binding interactions with FKBP, but is an intrinsic property of these precursors. Therefore, these molecules would be expected to copurify with other low-molecular-weight proteins, especially in the abbreviated purification protocols made possible by freeze-thaw release of recombinant proteins from E. coli (Johnson, B. H., and Hecht, M. H. (1994) BioTechnology 12, 1357-1360). Several alternative strategies are discussed for integrating these findings into the design of improved purification procedures for low-molecular-weight recombinant proteins.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Ram MK,Andrade LJ,Phillips TB,van Schravendijk MR

doi

10.1006/prep.1999.1145

keywords:

subject

Has Abstract

pub_date

1999-11-01 00:00:00

pages

305-11

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(99)91145-2

journal_volume

17

pub_type

杂志文章

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