EG10258

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Author Information

Selin Chiragzada

Basic Information

  • ID: EG10258
  • Name: eno (phosphopyruvate hydratase or enolase)
  • Organism: E. coli
  • Description:

Phosphopyruvate hydratase, or enolase, is an enzyme that catalyzes the interconversion of 2-phosphoglycerate and phosphoenolpyruvate. This occurs during the processes of gluconeogenesis and glycolysis (https://ecocyc.org/gene?orgid=ECOLI&id=ENOLASE-MONOMER). During glycolysis, enolase catalyzes the following reaction, which is also the penultimate step of the process:

2-phospho-D-glycerate <-> phosphoenolpyruvate + H2O

This is a 5-step reaction, and enolase is involved in the fourth step. Even though its role in RNA degradation is not fully known yet, it is has also shown to be a part of the RNA degradasome, a complex involved in RNA processing and degradation of messenger RNA. (http://www.uniprot.org/uniprot/P0A6P9). Enolase has a secondary structure and contains both alpha-helices and beta-pleated sheets. It is found in both the cytoplasm and the cell surface, where it forms part of the bacterial cytoskeleton. E.coli enolase is similar to other enolases (human, plant) in that it requires Mg2+ as a cofactor in the reaction and is inhibited by the fluoride ion in the presence of phosphate. Mg2+ binds to enolase, producing a change in its shape, so it is easier for 2-PGA to bind to it (http://proteopedia.org/wiki/index.php/Enolase). It is, however, different in that its optimum pH is higher than that of other enolases (https://ecocyc.org/gene?orgid=ECOLI&id=ENOLASE-MONOMER).

  • DNA Length: 1299 base pairs.
  • DNA sequence:

ATG AGC AAG ATC GTC AAA ATC ATT GGC AGA GAA ATC ATT GAT TCT CGT GGC AAC CCT ACA GTA GAG GCT GAG GTT CAT TTA GAG GGT GGC TTT GTC GGT ATG GCG GCG GCC CCG TCA GGT GCC TCT ACA GGT TCA CGG GAA GCG TTG GAG CTG CGT GAT GGC GAC AAG AGC AGA TTT TTA GGT AAG GGT GTT ACT AAG GCC GTA GCC GCT GTA AAT GGC CCT ATA GCG CAG GCG TTA ATC GGA AAG GAC GCC AAA GAT CAG GCG GGA ATA GAT AAG ATA ATG ATT GAC CTG GAT GGA ACT GAA AAC AAA AGC AAG TTC GGA GCT AAC GCA ATC TTA GCC GTC TCC CTG GCA AAC GCT AAA GCA GCG GCC GCG GCC AAA GGC ATG CCT CTG TAT GAG CAC ATA GCC GAA TTG AAC GGA ACA CCT GGA AAA TAT TCT ATG CCG GTG CCG ATG ATG AAC ATC ATT AAC GGA GGC GAG CAC GCG GAC AAT AAT GTG GAC ATA CAG GAG TTT ATG ATC CAG CCC GTC GGT GCG AAG ACG GTG AAA GAA GCA ATT CGC ATG GGA TCA GAG GTT TTC CAC CAT TTG GCT AAG GTT CTG AAG GCG AAA GGC ATG AAC ACT GCG GTT GGA GAC GAA GGG GGG TAT GCA CCT AAT CTT GGG TCG AAC GCA GAA GCG TTA GCG GTA ATT GCG GAA GCG GTA AAA GCT GCA GGG TAC GAG CTG GGA AAG GAT ATT ACA TTG GCA ATG GAT TGC GCC GCC TCA GAA TTT TAC AAG GAT GGG AAA TAT GTC TTA GCG GGA GAA GGT AAC AAG GCC TTT ACA TCG GAA GAA TTT ACT CAC TTC TTG GAA GAG TTG ACC AAA CAA TAT CCA ATT GTT AGT ATA GAA GAT GGA CTG GAC GAG TCA GAT TGG GAT GGG TTC GCT TAT CAA ACG AAA GTT CTG GGA GAT AAG ATA CAA TTG GTT GGT GAT GAC TTA TTC GTA ACG AAT ACC AAG ATA CTG AAA GAG GGA ATA GAA AAG GGA ATA GCC AAT TCG ATC CTT ATA AAG TTC AAC CAG ATC GGG TCT TTA ACT GAA ACA CTG GCT GCA ATT AAG ATG GCT AAA GAC GCT GGC TAT ACA GCG GTT ATT TCG CAC CGT TCT GGA GAG ACT GAG GAC GCC ACG ATA GCT GAC TTG GCC GTG GGC ACC GCT GCG GGA CAA ATT AAG ACC GGA AGC ATG TCG CGC TCC GAC CGG GTG GCG AAA TAT AAC CAG CTG ATT CGC ATT GAA GAG GCC CTG GGG GAG AAG GCA CCT TAT AAC GGC AGA AAG GAA ATT AAA GGA CAG GCT TAG

  • Amino Acid length: 432 amino acids.
  • Amino Acid sequence:

MSKIVKIIGR EIIDSRGNPT VEAEVHLEGG FVGMAAAPSG ASTGSREALE LRDGDKSRFL GKGVTKAVAA VNGPIAQALI GKDAKDQAGI DKIMIDLDGT ENKSKFGANA ILAVSLANAK AAAAAKGMPL YEHIAELNGT PGKYSMPVPM MNIINGGEHA DNNVDIQEFM IQPVGAKTVK EAIRMGSEVF HHLAKVLKAK GMNTAVGDEG GYAPNLGSNA EALAVIAEAV KAAGYELGKD ITLAMDCAAS EFYKDGKYVL AGEGNKAFTS EEFTHFLEEL TKQYPIVSIE DGLDESDWDG FAYQTKVLGD KIQLVGDDLF VTNTKILKEG IEKGIANSIL IKFNQIGSLT ETLAAIKMAK DAGYTAVISH RSGETEDATI ADLAVGTAAG QIKTGSMSRS DRVAKYNQLI RIEEALGEKA PYNGRKEIKG QA

Function and Homologs

  • Product: enolase enzyme
  • Module: glycolytic enzyme
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • MULTISPECIES: enolase [Proteobacteria], Max score = 871/Query Cover = 100% /E-Value = 0.00 /Ident = 100%, [WP_000036723.1]
    • phosphopyruvate hydratase [Escherichia albertii], Max score = 870/Query Cover = 100%/E-Value = 0.00 /Ident = 99%, [WP_059224450.1]
    • phosphopyruvate hydratase [Shigella boydii], Max score = 869/Query Cover = 100%/E-Value = 0.00 /Ident = 99%, [WP_073840385.1]
  • Equivalent E. coli / JCVI functional protein: MMSYN1 0213

Expression

  • Expression Level: high
  • Expression Level Hypothesis: Unlike the homologous JCVI gene, E.coli eno is highly expressed. This is because E.coli enolase has also shown to be a part of the RNA degradasome, a complex involved in RNA processing and degradation of messenger RNA. Since it has multiple functions in E.coli, its expression is high.
  • Expression Level References and Description: I used the E.coli proteome dataset (File:EcoliProteomicExpressionData.xlsx) and I'm sure about my choice since I was able to find the exact gene in the given database.
  • Expression Time: early
  • Expression Level Hypothesis: Enolase plays an important role in respiration, which is a vital process that occurs in all cells constantly. However, it would be more important for eno to be expressed early on in the development of the organism, so that it has a chance to grow.
  • Expression Time References and Description: I made this decision based on the protein's function & did not use any resources.

Gene Context

  • Other Components: EG12296
  • Possible Dependencies: enzymes, such as amylase, maltase and sucrase, that break down polysaccharide chains into glucose. Glucose is required for glycolysis to be able to occur.
  • Process: glycolysis (interconversion of 2-phosphoglycerate and phosphoenolpyruvate)

Construct

  • Synthesis Score: The synthesis score of your construct: 1, 2,3
  • Predicted Translation Rate: Prediction of construct translation rate from the RBS calculator
  • Design Notes and Details: For example, had to use a rare codon to fix folding energy;
  • GenBank File: A link to the GenBank file. file