The crystal structure of an engineered monomeric triosephosphate isomerase, monoTIM: the correct modelling of an eight-residue loop
- PMID: 16100954
- DOI: 10.1016/0969-2126(93)90021-8
The crystal structure of an engineered monomeric triosephosphate isomerase, monoTIM: the correct modelling of an eight-residue loop
Abstract
Background: The triosephosphate isomerase (TIM) fold is found in several different classes of enzymes, most of which are oligomers; TIM itself always functions as a very tight dimer. It has recently been shown that a monomeric form of TIM ('monoTIM') can be constructed by replacing a 15-residue interface loop, loop-3, with an eight-residue fragment; modelling suggests that this should result in a short strain-free turn, resulting in the subsequent helix, helix-A3, having an additional turn at its amino terminus.
Results: The crystal structure of monoTIM shows that it retains the characteristic TIM-barrel (betaalpha)8-fold and that the new loop has a structure very close to that predicted. Two other interface loops, loop-1 and loop-4, which contain the active site residues Lys13 and His95, respectively, show significant changes in structure in monoTIM compared with dimeric wild-type TIM.
Conclusion: The observed structural differences between monoTIM and wild-type TIM indicate that the dimeric appearance of TIM determines the location and conformation of two of the four catalytic residues.
Similar articles
-
Three new crystal structures of point mutation variants of monoTIM: conformational flexibility of loop-1, loop-4 and loop-8.Structure. 1995 Jul 15;3(7):669-79. doi: 10.1016/s0969-2126(01)00202-7. Structure. 1995. PMID: 8591044
-
Protein engineering with monomeric triosephosphate isomerase (monoTIM): the modelling and structure verification of a seven-residue loop.Protein Eng. 1997 Feb;10(2):159-67. doi: 10.1093/protein/10.2.159. Protein Eng. 1997. PMID: 9089815
-
Active site properties of monomeric triosephosphate isomerase (monoTIM) as deduced from mutational and structural studies.Protein Sci. 1996 Feb;5(2):229-39. doi: 10.1002/pro.5560050206. Protein Sci. 1996. PMID: 8745400 Free PMC article.
-
Triosephosphate isomerase: a highly evolved biocatalyst.Cell Mol Life Sci. 2010 Dec;67(23):3961-82. doi: 10.1007/s00018-010-0473-9. Epub 2010 Aug 7. Cell Mol Life Sci. 2010. PMID: 20694739 Free PMC article. Review.
-
The TIM-barrel fold: a versatile framework for efficient enzymes.FEBS Lett. 2001 Mar 16;492(3):193-8. doi: 10.1016/s0014-5793(01)02236-0. FEBS Lett. 2001. PMID: 11257493 Review.
Cited by
-
Crystal structures of two monomeric triosephosphate isomerase variants identified via a directed-evolution protocol selecting for L-arabinose isomerase activity.Acta Crystallogr F Struct Biol Commun. 2016 Jun;72(Pt 6):490-9. doi: 10.1107/S2053230X16007548. Epub 2016 May 23. Acta Crystallogr F Struct Biol Commun. 2016. PMID: 27303904 Free PMC article.
-
Identification of the Serratia endonuclease dimer: structural basis and implications for catalysis.Protein Sci. 1996 Jan;5(1):24-33. doi: 10.1002/pro.5560050104. Protein Sci. 1996. PMID: 8771193 Free PMC article.
-
Three-dimensional structure of 6-pyruvoyl tetrahydropterin synthase, an enzyme involved in tetrahydrobiopterin biosynthesis.EMBO J. 1994 Mar 15;13(6):1255-62. doi: 10.1002/j.1460-2075.1994.tb06377.x. EMBO J. 1994. PMID: 8137809 Free PMC article.
-
Dissection of the gene of the bifunctional PGK-TIM fusion protein from the hyperthermophilic bacterium Thermotoga maritima: design and characterization of the separate triosephosphate isomerase.Protein Sci. 1997 Oct;6(10):2159-65. doi: 10.1002/pro.5560061010. Protein Sci. 1997. PMID: 9336838 Free PMC article.
-
Insight into the functional roles of Glu175 in the hyperthermostable xylanase XYL10C-ΔN through structural analysis and site-saturation mutagenesis.Biotechnol Biofuels. 2018 Jun 8;11:159. doi: 10.1186/s13068-018-1150-8. eCollection 2018. Biotechnol Biofuels. 2018. PMID: 29930705 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources