MMSYN1 0216

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Author Information

Sharman Tan

Basic Information

  • ID: MMSYN1_0216
  • Name: HGPRT
  • Organism: Mycoplasma mycoides
  • Description: HGPRT is a purine salvage enzyme. A purine salvage pathway involves the synthesis of purine from intermediates in the degradative pathway for nucleotides. Salvage pathways are used to recover bases and nucleosides that are formed during degradation of RNA and DNA [1]. This is important in some organs because some tissues cannot undergo de novo synthesis [2]. In this case, HGPRT's primary activity involves hypoxanthine as the substrate. The enzyme's activity with guanine appears to be sufficient to salvage guanine for guanine nucleotide synthesis [3]. It is inactive towards xanthine [4]. The protein encoded by this gene is a transferase, which catalyzes conversion of hypoxanthine to inosine monophosphate and guanine to guanosine monophosphate via transfer of the 5-phosphoribosyl group from 5-phosphoribosyl 1-pyrophosphate. HGPRT is involved in a subpathway that is part of the IMP biosynthesis via salvage pathway, which is itself part of Purine metabolism [5].
  • DNA Length: 573 base pairs.
  • DNA sequence:

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

  • Amino Acid length: 190 amino acids.
  • Amino Acid sequence:

MQNLHPLVKEVLFTREQIQNRTKDIAKEIESYYKDKHLKDNSLLVVGLLKGCVPFYTDFCMVCDLTMEMDFMVVSSYHGSTSSNSAPKINLDLNTDVKDRDILIVEDIIDTGFTLKYVKEYLLNKGAKSVKILTMLDKPSGRKIDLVADWVCFTIDPCFVIGYGLDYQEKIRNLPYVAVCDTTKLDDWKW

Function and Homologs

  • Product: hypoxanthine phosphoribosyltransferase (HGPRTase)
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • hypoxanthine phosphoribosyltransferase [Mycoplasma capricolum], Max score: 373, Query Cover: 100%, E-value: 5e-131, Identity: 96%, WP_011387104.1
    • hypoxanthine phosphoribosyltransferase [Mycoplasma feriruminatoris], Max score: 361, Query Cover: 100%, E-value: 4e-126, Identity: 91%, WP_008363705.1
    • hypoxanthine phosphoribosyltransferase [Mycoplasma leachii], Max score: 349, Query Cover: 100%, E-value: 2e-121, Identity: 96%, WP_013447575.1
  • Equivalent E. coli / JCVI functional protein: EG20098.

Expression

  • Expression Level: Medium
  • Expression Level Hypothesis: Because HGPRT is a purine salvage enzyme, it is involved in the synthesis of purine from intermediates in the degradative pathway for nucleotides. Salvage pathways are used to recover bases and nucleosides that are formed during degradation of RNA and DNA, and since nucleotide synthesis is necessary for the organism, it makes sense that HGPRT has a medium expression level.
  • Expression Time: Right at the beginning
  • Expression Time Hypothesis: Because HGPRT is a purine salvage enzyme, it is involved in the synthesis of purine from intermediates in the degradative pathway for nucleotides. Salvage pathways are used to recover bases and nucleosides that are formed during degradation of RNA and DNA, and since nucleotide synthesis is immediately necessary for the organism, it makes sense that HGPRT is expressed right at the beginning of the life cycle of the organism.

Gene Context

  • Other Components: Because HPTGase is enough to catalyze the IMP biosynthesis via salvage pathway, no other components are necessary. [6]
  • Possible Dependencies: Because the inputs for the IMP biosynthesis via salvage pathway, hypoxanthine and guanine, is a naturally occurring purine derivative and a nucleic acid, respectively, there are no upstream enzymes that synthesize either of them. Therefore, there are no dependencies for HPTGase.
  • Process: IMP biosynthesis via salvage pathway
    • Inputs: hypoxanthine, guanine
    • Outputs: inosine monophosphate, guanosine monophosphate

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: [7].