Association of Heat-Induced Conformational Change with Activity Loss of Rubisco

Li, Guofu; Mao, Haibin; Ruan, Xiang; Xu, Qiang; Gong, Yu; Zhang, Xiufang; Zhao, Nanming

doi:10.1006/bbrc.2001.6322

Cited by 14 publications

(8 citation statements)

References 27 publications

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“…This behavior of the four proteins in this study is similar to other proteins that became inactive at temperatures that were lower than temperatures at which circular dicroism indicated unfolding28293031.…”

Section: Resultssupporting

confidence: 74%

Effect of arginine on oligomerization and stability of N-acetylglutamate synthase

Haskins

Mumo

Brown

et al. 2016

Sci Rep

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N-acetylglutamate synthase (NAGS; E.C.2.3.1.1) catalyzes the formation of N-acetylglutamate (NAG) from acetyl coenzyme A and glutamate. In microorganisms and plants, NAG is the first intermediate of the L-arginine biosynthesis; in animals, NAG is an allosteric activator of carbamylphosphate synthetase I and III. In some bacteria bifunctional N-acetylglutamate synthase-kinase (NAGS-K) catalyzes the first two steps of L-arginine biosynthesis. L-arginine inhibits NAGS in bacteria, fungi, and plants and activates NAGS in mammals. L-arginine increased thermal stability of the NAGS-K from Maricaulis maris (MmNAGS-K) while it destabilized the NAGS-K from Xanthomonas campestris (XcNAGS-K). Analytical gel chromatography and ultracentrifugation indicated tetrameric structure of the MmMNAGS-K in the presence and absence of L-arginine and a tetramer-octamer equilibrium that shifted towards tetramers upon binding of L-arginine for the XcNAGS-K. Analytical gel chromatography of mouse NAGS (mNAGS) indicated either different oligomerization states that are in moderate to slow exchange with each other or deviation from the spherical shape of the mNAGS protein. The partition coefficient of the mNAGS increased in the presence of L-arginine suggesting smaller hydrodynamic radius due to change in either conformation or oligomerization. Different effects of L-arginine on oligomerization of NAGS may have implications for efforts to determine the three-dimensional structure of mammalian NAGS.

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Section: Resultssupporting

confidence: 74%

Effect of arginine on oligomerization and stability of N-acetylglutamate synthase

Haskins

Mumo

Brown

et al. 2016

Sci Rep

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“…First, there is biochemical evidence that the thermal stability of RuBisCO is greater in more thermotolerant organisms. Although the loss of activity of spinach RuBisCO becomes partly irreversible above 45 °C (Li et al . 2002), enzyme activity is maximized at 55 °C in the thermophile Synechococcus strain Y52‐s (Sheridan & Ulik 1976).…”

Section: Introductionmentioning

confidence: 99%

Evidence for the adaptive evolution of the carbon fixation generbcLduring diversification in temperature tolerance of a clade of hot spring cyanobacteria

Miller

2003

Molecular Ecology

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Determining the molecular basis of enzyme adaptation is central to understanding the evolution of environmental tolerance but is complicated by the fact that not all amino acid differences between ecologically divergent taxa are adaptive. Analysing patterns of nucleotide sequence evolution can potentially guide the investigation of protein adaptation by identifying candidate codon sites on which diversifying selection has been operating. Here, I test whether there is evidence for molecular adaptation of the carbon fixation gene rbcL for a clade of hot spring cyanobacteria in the genus Synechococcus that has diverged in thermotolerance. Amino acid replacements during Synechococcus radiation have resulted in an increase in the number of hydrophobic residues in the RbcLs of more thermotolerant strains. A similar increase in hydrophobicity has been observed for many thermostable proteins. Maximum likelihood models which allow for heterogeneity among codon sites in the ratio of nonsynonymous to synonymous nucleotide substitutions estimated a class of amino acid sites as a target of positive selection. Depending on the model, a single amino acid site that interacts with a flexible element involved in the opening and closing of the active site was estimated with either low or moderate support to be a member of this class. Site-directed mutagenesis approaches are being explored in order to directly test its adaptive significance.

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“…For comparison, an IEC fraction of the corresponding extracts was also analysed (Additional file 4). Interestingly, each of the hydrophobically distinct fractions of Rubisco had similar values of molar ellipticites [θ] (~ -8000 deg cm 2 dmol -1 ) and structures of the CD spectra when compared to each other, to the IEC fraction, and with CD spectra of Rubisco previously reported in the literature [40,41]. For comparison, maximum denaturation of the fractions was achieved by heating at 80°C (shown in Figures 5B and 6B), which caused a large drop in the molar ellipticities of the samples (≤2000 deg cm 2 dmol -1 ), as well as a loss of structure in the spectra.…”

Section: Resultsmentioning

confidence: 86%

Isolation and kinetic characterisation of hydrophobically distinct populations of form I Rubisco

et al. 2014

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BackgroundRubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) is a Calvin Cycle enzyme involved in CO2 assimilation. It is thought to be a major cause of photosynthetic inefficiency, suffering from both a slow catalytic rate and lack of specificity due to a competing reaction with oxygen. Revealing and understanding the engineering rules that dictate Rubisco’s activity could have a significant impact on photosynthetic efficiency and crop yield.ResultsThis paper describes the purification and characterisation of a number of hydrophobically distinct populations of Rubisco from both Spinacia oleracea and Brassica oleracea extracts. The populations were obtained using a novel and rapid purification protocol that employs hydrophobic interaction chromatography (HIC) as a form I Rubisco enrichment procedure, resulting in distinct Rubisco populations of expected enzymatic activities, high purities and integrity.ConclusionsWe demonstrate here that HIC can be employed to isolate form I Rubisco with purities and activities comparable to those obtained via ion exchange chromatography (IEC). Interestingly, and in contrast to other published purification methods, HIC resulted in the isolation of a number of hydrophobically distinct Rubisco populations. Our findings reveal a so far unaccounted diversity in the hydrophobic properties within form 1 Rubisco. By employing HIC to isolate and characterise Spinacia oleracea and Brassica oleracea, we show that the presence of these distinct Rubisco populations is not species specific, and we report for the first time the kinetic properties of Rubisco from Brassica oleracea extracts. These observations may aid future studies concerning Rubisco’s structural and functional properties.

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Association of Heat-Induced Conformational Change with Activity Loss of Rubisco

Cited by 14 publications

References 27 publications

Effect of arginine on oligomerization and stability of N-acetylglutamate synthase

Effect of arginine on oligomerization and stability of N-acetylglutamate synthase

Evidence for the adaptive evolution of the carbon fixation generbcLduring diversification in temperature tolerance of a clade of hot spring cyanobacteria

Isolation and kinetic characterisation of hydrophobically distinct populations of form I Rubisco

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