Difference between revisions of "Part:BBa K4414006"

 
Line 1: Line 1:
 +
_NOTOC__
  
−
__NOTOC__
 
 
<partinfo>BBa_K4414006 short</partinfo>
 
<partinfo>BBa_K4414006 short</partinfo>
  
−
Report gene.The enzyme encoded by this gene catalyzes the first 2 steps, and at least 1 subsequent step, in the conversion of tyrosine to melanin.  
+
 
 +
 
 +
==Usage and Biology==
 +
 
 +
Tyrosinase（TYR） is a key enzyme in melanin synthesis that catalyzes the rate-limiting step of the reaction. By activating tyrosine kinase can increase the yield of melanin, forming melanin deposition .
 +
The melanogenesis process is initiated with the oxidation of L-tyrosine to dopaquinone (DQ) by TYR. The resulting quinone will serve as a substrate for the synthesis of eumelanin and pheomelanin(Hearing & Jiménez, 1987). The formation of DQ is a rate-limiting step in the melanin synthesis because remainder of the reaction sequence can proceed spontaneously at a physiological pH value(Halaban et al., 2002). After DQ formation, it undergoes intramolecular cyclization to produce indoline, leukodopachrome (cyclodopa). The redox exchange between leukodopachrome and DQ give rise to dopachrome and L-3,4-dihydroxyphenylalanine (L-DOPA), which is also a substrate for TYR and oxidized to DQ again by the enzyme. Dopachrome gradually decomposes to give dihydroxyindole (DHI) and dihydroxyindole-2-carboxylicacid (DHICA). The later process is catalyzed by TRP-2, now known as dopachrome tautomerase (DCT). Ultimately, these dihydroxyindoles (DHI and DHICA) are oxidized to eumelanin. TRP-1 is believed to catalyze the oxidation of DHICA to eumelanin. Alongside, DQ is converted to 5-S-cysteinyldopa or glutothionyldopa in the presence of cysteine or glutathione. Subsequent oxidation gives benzothiazine intermediates and finally to produce pheomelanin. Although three enzymes, TYR, TRP-1 and TRP-2 are involved in the melanogenesis pathway, tyrosinase is exclusively necessary for melanogenesis.(Pillaiyar et al., 2017)
  
  
  
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
  
 
<!-- -->
 
<!-- -->
−
<span class='h3bb'>Sequence and Features</span>
+
===Sequence and Features===
 
<partinfo>BBa_K4414006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4414006 SequenceAndFeatures</partinfo>
  
Line 19: Line 22:
 
<partinfo>BBa_K4414006 parameters</partinfo>
 
<partinfo>BBa_K4414006 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
 +
===Functional characterization and result===
 +
 +
We introduce the plasmid with TYR into the cells, where melanogen is produced as shown.
 +
 +
<figure class="figure">
 +
<img src="https://static.igem.wiki/teams/4414/wiki/017-3.png" class="figure-img img-fluid rounded"  height="250px">
 +
 +
</figure>
 +
 +
</html>
 +
 +
Figure 1: (A)The blackening of HEK-293T cells at 24 h or 48 h post glucocorticoids treatment in microscope.(B)The blackening of HEK-293T cells at 24 h or 48 h post glucocorticoids treatment under the naked eye.
 +
 +
 +
===Reference===
 +
[1].Weikum ER, Knuesel MT, Ortlund EA, Yamamoto KR. Glucocorticoid receptor control of transcription: precision and plasticity via allostery. Nat Rev Mol Cell Biol. 2017 Mar;18(3):159-174. doi: 10.1038/nrm.2016.152. Epub 2017 Jan 5. PMID: 28053348; PMCID: PMC6257982.

Revision as of 11:21, 9 October 2022

_NOTOC__

TYR


Usage and Biology

Tyrosinase（TYR） is a key enzyme in melanin synthesis that catalyzes the rate-limiting step of the reaction. By activating tyrosine kinase can increase the yield of melanin, forming melanin deposition . The melanogenesis process is initiated with the oxidation of L-tyrosine to dopaquinone (DQ) by TYR. The resulting quinone will serve as a substrate for the synthesis of eumelanin and pheomelanin(Hearing & Jiménez, 1987). The formation of DQ is a rate-limiting step in the melanin synthesis because remainder of the reaction sequence can proceed spontaneously at a physiological pH value(Halaban et al., 2002). After DQ formation, it undergoes intramolecular cyclization to produce indoline, leukodopachrome (cyclodopa). The redox exchange between leukodopachrome and DQ give rise to dopachrome and L-3,4-dihydroxyphenylalanine (L-DOPA), which is also a substrate for TYR and oxidized to DQ again by the enzyme. Dopachrome gradually decomposes to give dihydroxyindole (DHI) and dihydroxyindole-2-carboxylicacid (DHICA). The later process is catalyzed by TRP-2, now known as dopachrome tautomerase (DCT). Ultimately, these dihydroxyindoles (DHI and DHICA) are oxidized to eumelanin. TRP-1 is believed to catalyze the oxidation of DHICA to eumelanin. Alongside, DQ is converted to 5-S-cysteinyldopa or glutothionyldopa in the presence of cysteine or glutathione. Subsequent oxidation gives benzothiazine intermediates and finally to produce pheomelanin. Although three enzymes, TYR, TRP-1 and TRP-2 are involved in the melanogenesis pathway, tyrosinase is exclusively necessary for melanogenesis.(Pillaiyar et al., 2017)



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Functional characterization and result

We introduce the plasmid with TYR into the cells, where melanogen is produced as shown.

<figure class="figure"> <img src="017-3.png" class="figure-img img-fluid rounded" height="250px">

</figure>

</html>

Figure 1: (A)The blackening of HEK-293T cells at 24 h or 48 h post glucocorticoids treatment in microscope.(B)The blackening of HEK-293T cells at 24 h or 48 h post glucocorticoids treatment under the naked eye.


Reference

[1].Weikum ER, Knuesel MT, Ortlund EA, Yamamoto KR. Glucocorticoid receptor control of transcription: precision and plasticity via allostery. Nat Rev Mol Cell Biol. 2017 Mar;18(3):159-174. doi: 10.1038/nrm.2016.152. Epub 2017 Jan 5. PMID: 28053348; PMCID: PMC6257982.