The Protein Journal

, Volume 37, Issue 5, pp 407–427 | Cite as

Nonfunctional Missense Mutants in Two Well Characterized Cytosolic Enzymes Reveal Important Information About Protein Structure and Function

  • Ashley E. Cole
  • Fatmah M. Hani
  • Brian W. Allen
  • Paul C. Kline
  • Elliot AltmanEmail author


The isolation and characterization of 42 unique nonfunctional missense mutants in the bacterial cytosolic β-galactosidase and catechol 2,3-dioxygenase enzymes allowed us to examine some of the basic general trends regarding protein structure and function. A total of 6 out of the 42, or 14.29% of the missense mutants were in α-helices, 17 out of the 42, or 40.48%, of the missense mutants were in β-sheets and 19 out of the 42, or 45.24% of the missense mutants were in unstructured coil, turn or loop regions. While α-helices and β-sheets are undeniably important in protein structure, our results clearly indicate that the unstructured regions are just as important. A total of 21 out of the 42, or 50.00% of the missense mutants caused either amino acids located on the surface of the protein to shift from hydrophilic to hydrophobic or buried amino acids to shift from hydrophobic to hydrophilic and resulted in drastic changes in hydropathy that would not be preferable. There was generally good consensus amongst the widely used algorithms, Chou–Fasman, GOR, Qian–Sejnowski, JPred, PSIPRED, Porter and SPIDER, in their ability to predict the presence of the secondary structures that were affected by the missense mutants and most of the algorithms predicted that the majority of the 42 inactive missense mutants would impact the α-helical and β-sheet secondary structures or the unstructured coil, turn or loop regions that they altered.


Protein secondary structure α-Helices β-Sheets Unstructured regions Coils Hydropathy 


Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Authors and Affiliations

  1. 1.Department of BiologyMiddle Tennessee State UniversityMurfreesboroUSA
  2. 2.Department of ChemistryMiddle Tennessee State UniversityMurfreesboroUSA
  3. 3.Department of Biomedical EngineeringDuke UniversityDurhamUSA

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