Determination of the amino acid requirements for a protein hinge in triosephosphate isomerase
| Autor: | Nicole S. Sampson, Jeonghoon Sun |
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| Rok vydání: | 1998 |
| Předmět: |
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Molecular Protein Conformation Molecular Sequence Data Mutant Sequence alignment Biology Glyceraldehyde 3-Phosphate Biochemistry Catalysis Triosephosphate isomerase Structure-Activity Relationship Protein structure Animals Amino Acid Sequence Molecular Biology Peptide sequence Gene Library Genetics Binding Sites Base Sequence Genetic Complementation Test Sequence Analysis DNA Cassette mutagenesis Mutagenesis Insertional Amino Acid Substitution Sequence space (evolution) Genetic library Chickens Sequence Alignment Triose-Phosphate Isomerase Research Article |
| Zdroj: | Protein Science. 7:1495-1505 |
| ISSN: | 1469-896X 0961-8368 |
| DOI: | 10.1002/pro.5560070702 |
| Popis: | We have determined the sequence requirements for a protein hinge in triosephosphate isomerase. The codons encoding the hinge at the C-terminus of the active-site lid of triosephosphate isomerase were replaced with a genetic library of all possible 8,000 amino acid combinations. The most active of these 8,000 mutants were selected using in vivo complementation of a triosephosphate isomerase deficient strain of E. coli, DF502. Approximately 3% of the mutants complement DF502 with an activity that is above 70% of wild-type activity. The sequences of these hinge mutants reveal that the solutions to the hinge flexibility problem are varied. Moreover, these preferences are sequence dependent; that is, certain pairs occur frequently. They fall into six families of similar sequences. In addition to the hinge sequences expected on the basis of phylogenetic analysis, we selected three new families of 3-amino-acid hinges: X(A/S)(L/K/M), X(aromatic/beta-branched)(L/K), and XP(S/N). The absence of these hinge families in the more than 60 known species of triosephosphate isomerase suggests that during evolution, not all of sequence space is sampled, perhaps because there is no neutral mutation pathway to access the other families. |
| Databáze: | OpenAIRE |
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