Figures & data
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Fig. 1. Phylogenetic tree and TM topological model of AGCS family proteins.
Notes: (A) Phylogenetic tree of ApAgcS1, ApAgcS2, Acp from thermophilic bacterium PS3, Pseudomonas AgcS, and DagA from A. haloplanktis. The tree was constructed from alignments of full-length amino acid sequences using the Clustal W and TreeView programs. (B) TM topology model of ApAgcS1. TM topology model of ApAgcS1 was calculated by an algorithm TMHMM.
![Fig. 1. Phylogenetic tree and TM topological model of AGCS family proteins.Notes: (A) Phylogenetic tree of ApAgcS1, ApAgcS2, Acp from thermophilic bacterium PS3, Pseudomonas AgcS, and DagA from A. haloplanktis. The tree was constructed from alignments of full-length amino acid sequences using the Clustal W and TreeView programs. (B) TM topology model of ApAgcS1. TM topology model of ApAgcS1 was calculated by an algorithm TMHMM.](/cms/asset/d78adbe7-2a34-4d13-ad96-4b3ee1a7b1f9/tbbb_a_968091_f0001_oc.gif)
Fig. 2. Uptake of glycine by ApAgcS1.
Notes: (A) Time course of glycine uptake by ApAgcS1. B) Effects of NaCl, KCl, and LiCl on glycine uptake by ApAgcS1. ApAgcS1 expressing E. coli JW4166 cells were grown at 37 °C as described in “Materials and Methods.” Glycine uptake was measured after 10 μM [U-14C] glycine was added. In (B), duration of uptake was 1 min. Each value represents the average of three independent measurements and error bars represent standard deviations.
![Fig. 2. Uptake of glycine by ApAgcS1.Notes: (A) Time course of glycine uptake by ApAgcS1. B) Effects of NaCl, KCl, and LiCl on glycine uptake by ApAgcS1. ApAgcS1 expressing E. coli JW4166 cells were grown at 37 °C as described in “Materials and Methods.” Glycine uptake was measured after 10 μM [U-14C] glycine was added. In (B), duration of uptake was 1 min. Each value represents the average of three independent measurements and error bars represent standard deviations.](/cms/asset/86b49372-2247-407c-aea2-f6c3991d03ac/tbbb_a_968091_f0002_b.gif)
Fig. 3. Kinetics of glycine uptake by ApAgcS1.
Notes: (A) Saturation curve. (B) Effects of pH. Glycine uptake by ApAgcS1 in JW4166 cells was assayed in the presence of 0.3 M NaCl. The pH for saturation curve was 7.4. Duration of uptake was 1 min. Each value represents the average of three independent measurements and error bars represent standard deviations.
![Fig. 3. Kinetics of glycine uptake by ApAgcS1.Notes: (A) Saturation curve. (B) Effects of pH. Glycine uptake by ApAgcS1 in JW4166 cells was assayed in the presence of 0.3 M NaCl. The pH for saturation curve was 7.4. Duration of uptake was 1 min. Each value represents the average of three independent measurements and error bars represent standard deviations.](/cms/asset/8f26afe1-82d8-488c-9a4f-9629f21598f8/tbbb_a_968091_f0003_b.gif)
Fig. 4. Competition of glycine uptake by ApAgcS1.
Notes: The reaction mixtures contained 1 μM [U-14C]-glycine and 0.3 M NaCl. The concentration of various competitors was 100 μM. The value of uptake in the control (C) without a competitor is shown as 100%. Each value represents the average of three independent measurements and error bars represent standard deviations.
![Fig. 4. Competition of glycine uptake by ApAgcS1.Notes: The reaction mixtures contained 1 μM [U-14C]-glycine and 0.3 M NaCl. The concentration of various competitors was 100 μM. The value of uptake in the control (C) without a competitor is shown as 100%. Each value represents the average of three independent measurements and error bars represent standard deviations.](/cms/asset/e28162fe-809c-4cb6-a81a-39f814e90ef3/tbbb_a_968091_f0004_b.gif)
Fig. 5. Expression of mRNA for ApagcS1.
Notes: (A) Nitrate-deficient stress. (B) Salt stress (2.5 M NaCl). A. halophytica cells were collected after 0, 1, 3, 6, 12, and 24 h growth. RT-PCR analysis was performed as described in “Materials and Methods.” The ribonuclease P(rnpB) gene was used as the control. The values at time 0 for each gene were set to 1.0. Asterisk indicates significant difference (p < 0.05) from the values at time 0. Each value represents the average of three independent measurements and error bars represent standard deviations.
![Fig. 5. Expression of mRNA for ApagcS1.Notes: (A) Nitrate-deficient stress. (B) Salt stress (2.5 M NaCl). A. halophytica cells were collected after 0, 1, 3, 6, 12, and 24 h growth. RT-PCR analysis was performed as described in “Materials and Methods.” The ribonuclease P(rnpB) gene was used as the control. The values at time 0 for each gene were set to 1.0. Asterisk indicates significant difference (p < 0.05) from the values at time 0. Each value represents the average of three independent measurements and error bars represent standard deviations.](/cms/asset/9deab5ee-45fe-4bc4-8801-8078c574b95a/tbbb_a_968091_f0005_oc.gif)
Fig. 6. Extracellular amino acid accumulation in the culture medium.
Notes: The extracellular amino acids in the growth medium after 24 h incubation of S. elongatus PCC 7942 were measured. Wild-type, gray bar; ΔnatG mutant harbored empty vector, black bar; ΔnatG mutant harbored ApagcS1, white bar. Asterisk indicates significant difference (p < 0.05) between S. elongatus PCC 7942 cells expressing natG_empty vector_24 h and natG_pSyn1_ApagcS1_24 h. Each value represents the average of three independent measurements and error bars represent standard deviations.
![Fig. 6. Extracellular amino acid accumulation in the culture medium.Notes: The extracellular amino acids in the growth medium after 24 h incubation of S. elongatus PCC 7942 were measured. Wild-type, gray bar; ΔnatG mutant harbored empty vector, black bar; ΔnatG mutant harbored ApagcS1, white bar. Asterisk indicates significant difference (p < 0.05) between S. elongatus PCC 7942 cells expressing natG_empty vector_24 h and natG_pSyn1_ApagcS1_24 h. Each value represents the average of three independent measurements and error bars represent standard deviations.](/cms/asset/0712c20a-357b-4f07-a754-26963dd40330/tbbb_a_968091_f0006_b.gif)