Mapping tertiary interactions in protein folding reactions: a novel mass spectrometry- and chemical synthesis-based approach.


Journal Article

A novel mass spectrometry- and chemical synthesis-based approach for studying protein folding reactions is described, and its initial application to study the folding/unfolding reaction of a homo-hexameric enzyme 4-oxalocrotonate (4OT) is reported. This new approach involves the application of total chemical synthesis to prepare protein analogues that contain a photoreactive amino acid site-specifically incorporated into their primary amino acid sequence. To this end, a photoreactive amino acid-containing analogue of 4OT in which Pro-1 was replaced with p-benzoyl-l-phenylalanine (Bpa) was prepared. This analogue can be used to map structurally specific protein-protein interactions in 4OT's native folded state. These photocrosslinking studies and peptide mapping results with (PlBpa)4OT indicate that this construct is potentially useful for probing the structural properties of equilibrium and kinetic intermediates in 4OT's folding reaction.

Full Text

Duke Authors

Cited Authors

  • Moss, JA; Silinski, P; Fitzgerald, MC

Published Date

  • February 2001

Published In

Volume / Issue

  • 369 / 3-4

Start / End Page

  • 252 - 257

PubMed ID

  • 11293701

Pubmed Central ID

  • 11293701

Electronic International Standard Serial Number (EISSN)

  • 1432-1130

International Standard Serial Number (ISSN)

  • 0937-0633

Digital Object Identifier (DOI)

  • 10.1007/s002160000644


  • eng