Part:BBa_K4620016
Skr-BURP tryptic peptide with Y- deletion containing the core peptide
ADNVLFPSYAAR is a modification of ADNVLFYPSYAAR, a peptide formed after tryptic digestion of Skr-BURP protein. It contains core peptide fragment VLFYPSY. VLFYPSY is a peptide that can be cyclised by BURP domain protein from African clubmoss (Selaginella kraussiana) (BBa_K460000), forming selanine A. This side chain macrocyclisation reaction is catalysed by copper (II) ions. Cyclisation happens between the OH-group of tyrosine and Cβ of leucine or Cβ of tyrosine. A bicyclic compound is formed. As of this day, mechanism of this reaction has not been fully characterised.
<"
">
In the nature, ADNVLFYPSYAAR is conjugated to the N-terminal domain of Skr-BURP. iGEM 2023 team Latvia-Riga produced this peptide separately in order to make trans-cyclisation reaction where the cyclisation substrate is not attached to the catalytic domain. We chose substrate a little bit longer than the core domain in order to make expression and purification easier.
We designed several modificated core peptide analogues in order to see what are the limits of SkrBURP cyclisation capacity. In this peptide we removed one tyrosine residue to see if the cycle would still be formed. Our team cloned ADNVLFPSYAAR gene into pEXP-GB1 expression vector, containing GB1 (domain B1 of Immunoglobulin G-binding protein G from Streptococcus sp.), N-terminal 8xHis tag and TEV protease cleavage site.
<"
">
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |