Molecular Determinants for Protein Stabilization by Insertional Fusion to a Thermophilic Host Protein

Brennal Pierre, Jason W. Labonte, Tina Xiong, Edwin Aoraha, Asher Williams, Vandan Shah, Edward Chau, Kazi Yasin Helal, Jeffrey J. Gray, Jin Ryoun Kim

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


A universal method that improves protein stability and evolution has thus far eluded discovery. Recently, however, studies have shown that insertional fusion to a protein chaperone stabilized various target proteins with minimal negative effects. The improved stability was derived from insertion into a hyperthermophilic protein, Pyrococcus furiosus maltodextrin-binding protein (PfMBP), rather than from changes to the target protein sequence. In this report, by evaluating the thermodynamic and kinetic stability of various inserted β-lactamase (BLA) homologues, we were able to examine the molecular determinants of stability realized by insertional fusion to PfMBP. Results indicated that enhanced stability and suppressed aggregation of BLA stemmed from enthalpic and entropic mechanisms. This report also suggests that insertional fusion to a stable protein scaffold has the potential to be a useful method for improving protein stability, as well as functional protein evolution.

Original languageEnglish (US)
Pages (from-to)2392-2402
Number of pages11
Issue number16
StatePublished - Nov 1 2015


  • protein aggregation
  • protein engineering
  • protein fusion
  • protein stability
  • protein-protein interactions

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry


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