PMID:22037946

Citation	Zhang, Y, Gao, X, Zheng, Y and Garavito, RM (2011) Identification of succinic semialdehyde reductases from Geobacter: expression, purification, crystallization, preliminary functional, and crystallographic analysis. Acta Biochim. Biophys. Sin. (Shanghai) 43:996-1002
Abstract	Succinic semialdehyde reductase (SSAR) is an important enzyme involved in γ-aminobutyrate (GABA) metabolism. By converting succinic semialdehyde (SSA) to γ-hydroxybutyrate (GHB), the SSAR facilitates an alternative pathway for GABA degradation. In this study, we identified SSARs from Geobacter sulfurreducens and Geobacter metallireducens (GsSSAR and GmSSAR, respectively). The enzymes were over-expressed in Escherichia coli and purified to near homogeneity. Both GsSSAR and GmSSAR showed the activity of reducing SSA using nicotinamide adenine dinucleotide phosphate as a co-factor. The oligomeric sizes of GsSSAR and GmSSAR, as determined by analytical size exclusion chromatography, suggest that the enzymes presumably exist as tetramers in solution. The recombinant GsSSAR and GmSSAR crystallized in the presence of NADP(+), and the resulting crystals diffracted to 1.89 Å (GsSSAR) and 2.25 Å (GmSSAR) resolution. The GsSSAR and GmSSAR crystals belong to the space groups P2(1)22(1) (a= 99.61 Å, b= 147.49 Å, c= 182.47 Å) and P1 (a= 75.97 Å, b= 79.14 Å, c= 95.47 Å, α = 82.15°, β = 88.80°, γ = 87.66°), respectively. Preliminary crystallographic data analysis suggests the presence of eight protein monomers in the asymmetric units for both GsSSAR and GmSSAR.
Links	PubMed Online version:10.1093/abbs/gmr097
Keywords	Amino Acid Sequence; Chromatography, Ion Exchange/methods; Crystallization/methods; Crystallography, X-Ray/methods; Geobacter/enzymology; Geobacter/metabolism; Hydroxybutyrate Dehydrogenase/chemistry; Hydroxybutyrate Dehydrogenase/genetics; Hydroxybutyrate Dehydrogenase/isolation & purification; Molecular Sequence Data; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
GEOMG:Q39Q67		GO:0009013: succinate-semialdehyde dehydrogenase [NAD(P)+] activity	ECO:0000314: direct assay evidence used in manual assertion		F		Figure 3 shows the succinic semialdehyde reducing ability to reducing SSA and demonstrated that NADPH/NADP+ is the suitable co-factor for these enzymes.	complete CACAO 8978
GEOMG:Q39Q67		GO:0009013: succinate-semialdehyde dehydrogenase [NAD(P)+] activity	ECO:0000247: sequence alignment evidence used in manual assertion	PMID:10320398	F		Table 4 shows the similarities in sequence between the SSAR proteins from Geobacter sulfurreducens and Geobacter metallireducens as compared to Arabidopsis thaliana, showing 64% sequence similarity	complete CACAO 8979

References

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