Molecular Dynamics Analysis of the Structural and Dynamic Properties of the Functionally Enhanced Hepta-variant of Mouse 5-aminolevulinate Synthase
Document Type
Article
Publication Date
2018
Keywords
5-aminolevulinate synthase, molecular dynamic, active site loop
Digital Object Identifier (DOI)
https://doi.org/10.1080/07391102.2016.1269688
Abstract
Heme biosynthesis, a complex, multistage, and tightly controlled process, starts with 5-aminolevulinate (ALA) production, which, in metazoa and certain bacteria, is a reaction catalyzed by 5-aminolevulinate synthase (ALAS), a pyridoxal 5′-phosphate (PLP)-dependent enzyme. Functional aberrations in ALAS are associated with several human diseases. ALAS can adopt open and closed conformations, with segmental rearrangements of a C-terminal, 16-amino acid loop and an α-helix regulating accessibility to the ALAS active site. Of the murine erythroid ALAS (mALAS2) forms previously engineered to assess the role of the flexible C-terminal loop versus mALAS2 function one stood out due to its impressive gain in catalytic power. To elucidate how the simultaneously introduced seven mutations of this activity-enhanced variant affected structural and dynamic properties of mALAS2, we conducted extensive molecular dynamics simulation analysis of the dimeric forms of wild-type mALAS2, hepta-variant and Rhodobacter capsulatus ALAS (aka R. capsulatus HemA). This analysis revealed that the seven simultaneous mutations in the C-terminal loop, which extends over the active site of the enzyme, caused the bacterial and murine proteins to adopt different conformations. Specifically, a new β-strand in the mutated ‘loop’ led to interaction with two preexisting β-strands and formation of an anti-parallel three-stranded β-sheet, which likely endowed the murine hepta-variant a more ‘stable’ open conformation than that of wild-type mALAS2, consistent with a kinetic mechanism involving a faster closed-to-open conformation transition and product release for the mutated than wild-type enzyme. Further, the dynamic behavior of the mALAS2 protomers was strikingly different in the two dimeric forms.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Journal of Biomolecular Structure and Dynamics, v. 36, issue 1, p. 152-165
Scholar Commons Citation
Na, Insung; DeForte, Shelly; Stojanovski, Bosko M.; Ferreira, Gloria C.; and Uversky, Vladimir N., "Molecular Dynamics Analysis of the Structural and Dynamic Properties of the Functionally Enhanced Hepta-variant of Mouse 5-aminolevulinate Synthase" (2018). Molecular Medicine Faculty Publications. 229.
https://digitalcommons.usf.edu/mme_facpub/229
