MMSYN1 0213

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Author Information

Selin Chiragzada

Basic Information

  • ID: MMSYN1_0213
  • Name: eno (phosphopyruvate hydratase or enolase)
  • Organism: JCVI_Syn.3.0
  • 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 (http://www.uniprot.org/uniprot/F4MPI1), both of which are important for the cell to survive. Glycolysis is a part of cellular respiration, so cells cannot survive without it. 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. 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. Enolase 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).

  • DNA Length: 1356 base pairs.
  • DNA sequence:

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

  • Amino Acid length: 451 amino acids.
  • Amino Acid sequence:

MSRIERIFAREILDSRGTPTVEVEVWTEFGGYGCAKAPSGASTGVNEALELRDGDKARYNGKGVLKAVKNVNEIIAPKLIGIDALDQLTVDRIMLDLDGTEFKTKLGANGILAVSLAVAKAAASELDIPLYKYLGGVQAKKLPVPMLNVINGGEHADSAIDFQEFMIMPVGAKSFSEALRWSSETFQALKSLLKSKKDITAVGDEGGFAPNFEWAYEKHDLKSFKAKTPAEIALDLLVEAIKKAGYKPGKDGIMIAMDCASSELYLEDKKYHFKKIEKVTNQEWSLTTDEMISYLEKLVDNYPIISIEDGLAETDWEGFTKLTQKIGDKVQIVGDDLFTTNPKFIKQGISKKAANSTLIKLNQIGTLSETVEAITMTQKAGWTAVVSHRSGETEDTTIADLAVAFNTGQIKTGSMSRSDRIAKYNRLLQIESELEKNAVYDGLEAFYNLNK

Function and Homologs

  • Functional Category: Glucose transport & catabolism
  • Product: phosphopyruvate hydratase enzyme
  • Module: glycolytic pathway
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • phosphopyruvate hydratase [Mycoplasma mycoides subsp. mycoides], Max score = 909/Query Cover = 99%/E-Value = 0.00 /Ident = 99%, [AME13526.1]
    • phosphopyruvate hydratase [Mycoplasma leachii], Max score = 907/Query Cover = 100%/E-Value = 0.00 /Ident = 98%, [WP_013447573.1]
    • enolase [Mycoplasma capricolum], Max score = 900/Query Cover = 100%/E-Value = 0.00 /Ident = 98%, [WP_011387101.1]
  • Equivalent E. coli / JCVI functional protein: EG10258.

Expression

  • Expression Level: low
  • Expression Level Hypothesis: Although enolase plays an important role in cell metabolism, it has only 1 function i.e. the catalysis of the interconversion of 2-phosphoglycerate and phosphoenolpyruvate. Therefore, the cell does not need to produce a lot of enolase.
  • Expression Level References and Description: I used the Mycoplasma genitalium database (File:MgenitaliumSimProteinCounts.xlsx). I am sure about my choice because I found the exact same protein in Mycoplasma genitalium, so their expression levels must be the same.
  • 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: pgm_bpd_ind: phosphoglycerate mutase MMSYN1 0729
  • 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