Skip to main content

Use of differential scanning fluorimetry to optimize the purification and crystallization of PLP‐dependent enzymes

Buy Article:

$43.00 plus tax (Refund Policy)

Differential scanning fluorimetry (DSF) is a practical and accessible technique that allows the assessment of multiphasic unfolding behavior resulting from subsaturating binding of ligands. Multiphasic unfolding is indicative of a heterogenous protein solution, which frequently interferes with crystallization and complicates functional characterization of proteins of interest. Along with UV–Vis spectroscopy, DSF was used to guide purification and crystallization improvements for the pyridoxal 5′‐phosphate (PLP) dependent transaminase BioA from Mycobacterium tuberculosis. The incompatibility of the primary amine‐containing buffer 2‐amino‐2‐(hydroxymethyl)‐1,3‐propanediol (Tris) and PLP was identified as a significant contributor to heterogeneity. It is likely that the utility of DSF for ligand‐binding assessment is not limited to the cofactor PLP but will be applicable to a variety of ligand‐dependent enzymes.
No References
No Citations
No Supplementary Data
No Article Media
No Metrics

Document Type: Research Article

Affiliations: Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA

Publication date: 2012-05-01

  • Access Key
  • Free content
  • Partial Free content
  • New content
  • Open access content
  • Partial Open access content
  • Subscribed content
  • Partial Subscribed content
  • Free trial content
Cookie Policy
X
Cookie Policy
Ingenta Connect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more