Using a residue clash map to functionally characterize protein recombination hybrids

Manish C. Saraf, Costas D. Maranas

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

In this article, we introduce a rapid, protein sequence database-driven approach to characterize all contacting residue pairs present in protein hybrids for inconsistency with protein family structural features. This approach is based on examining contacting residue pairs with different parental origins for different types of potentially unfavorable interactions (i.e. electrostatic repulsion, steric hindrance, cavity formation and hydrogen bond disruption). The identified clashing residue pairs between members of a protein family are then contrasted against functionally characterized hybrid libraries. Comparisons for five different protein recombination studies available in the literature: (i) glycinamide ribonucleotide transformylase (GART) from Escherichia coli (purN) and human (hGART), (ii) human Mu class glutathione S-transferase (GST) M1-1 and M2-2, (iii) β-lactamase TEM-1 and PSE-4, (iv) catechol-2, 3-oxygenase xylE and nahH, and (v) dioxygenases (toluene dioxygenase, tetrachlorobenzene dioxygenase and biphenyl dioxygenase) reveal that the patterns of identified clashing residue pairs are remarkably consistent with experimentally found patterns of functional crossover profiles. Specifically, we show that the proposed residue clash maps are on average 5.0 times more effective than randomly generated clashes and 1.6 times more effective than residue contact maps at explaining the observed crossover distributions among functional members of hybrid libraries. This suggests that residue clash maps can provide quantitative guidelines for the placement of crossovers in the design of protein recombination experiments.

Original languageEnglish (US)
Pages (from-to)1025-1034
Number of pages10
JournalProtein Engineering
Volume16
Issue number12
DOIs
StatePublished - Dec 2003

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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