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PMID:24358297
| Citation |
Rivera-Ordaz, A, Bracher, S, Sarrach, S, Li, Z, Shi, L, Quick, M, Hilger, D, Haas, R and Jung, H (2013) The sodium/proline transporter PutP of Helicobacter pylori. PLoS ONE 8:e83576 |
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| Abstract |
Helicobacter pylori is cause of chronic gastritis, duodenal ulcer and gastric carcinoma in humans. L-proline is a preferred energy source of the microaerophilic bacterium. Previous analyses revealed that HpputP and HpputA, the genes that are predicted to play a central role in proline metabolism as they encode for the proline transporter and proline dehydrogenase, respectively, are essential for stomach colonization. Here, the molecular basis of proline transport in H. pylori by HpPutP was investigated experimentally for the first time. Measuring radiolabeled substrate transport in H. pylori and E. coli heterologously expressing HpputP as well as in proteoliposomes reconstituted with HpPutP, we demonstrate that the observed proline transport in H. pylori is mediated by HpPutP. HpPutP is specific and exhibits a high affinity for L-proline. Notably, L-proline transport is exclusively dependent on Na(+) as coupling ion, i.e., Na(+)/L-proline symport, reminiscent to the properties of PutP of E. coli even though H. pylori lives in a more acidic environment. Homology model-based structural comparisons and substitution analyses identified amino acids crucial for function. HpPutP-catalyzed proline uptake was efficiently inhibited by the known proline analogs 3,4-dehydro-D,L-proline and L-azetidine-2-carboxylic acid. |
| Links |
PubMed PMC3866251 Online version:10.1371/journal.pone.0083576 |
| Keywords |
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| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0005296: L-proline permease activity |
ECO:0000314: |
F |
Figure 2a: A direct assay is preformed upon the H. pylori gene, HpPutP, in E coli. The uptake of L-proline into E. coli (harboring HpPutP) was much greater than cells transformed with pT7-5, the negative control in this experiment. Without HpPutP present in the cell, L-proline cannot be transported into the cell. |
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