Part:BBa_K4620015
Skr-BURP tryptic peptide containing the core peptide
ADNVLFYPSYAAR is 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_K4620000), 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. 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.
Cloning
Our team cloned ADNVLFYPSYAAR 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 |