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OaAEP1b-C247A

Host Organism

Oldenlandia affinis (Roem. & Schult.) DC.

Type

PAL

Sequence

MVRYLAGAVLLLVVLSVAAAVSGARDGDYLHLPSEVSRFFRPQETNDDHGEDSVGTRWAVLIAGSKGYANYRHQAGVCHAYQILKRGGLKDENIVVFMYDDIAYNESNPRPGVIINSPHGSDVYAGVPKDYTGEEVNAKNFLAAILGNKSAITGGSGKVVDSGPNDHIFIYYTDHGAAGVIGMPSKPYLYADELNDALKKKHASGTYKSLVFYLEACESGSMFEGILPEDLNIYALTSTNTTESSWAYYCPAQENPPPPEYNVCLGDLFSVAWLEDSDVQNSWYETLNQQYHHVDKRISHASHATQYGNLKLGEEGLFVYMGSNPANDNYTSLDGNALTPSSIVVNQRDADLLHLWEKFRKAPEGSARKEEAQTQIFKAMSHRVHIDSSIKLIGKLLFGIEKCTEILNAVRPAGQPLVDDWACLRSLVGTFETHCGSLSEYGMRHTRTIANICNAGISEEQMAEAASQACASIP

External Links

PDB ID: -
EC: 3.4.22.34
MEROPS: C13 legumain
UniProt ID: A0A0N9JZ32
NCBI: -

Substrate Specificity

P1
P1'
P2'
N/D
X
L/I

Functions

  • Intermolecular backbone terminal ligation
  • Backbone terminal cyclization
  • Side-chain to C-terminal cyclization
  • Specificity stringency: Prime-low, Nonprime-high

Expression Platform:

  • E. coli
  • N. benthamiana

Mutations:

  • Fast kinetics: C247A

Other Properties:

-

References:

  1. [1] (Engineering, Structure) Yang, R. L.; Wong, Y. H.; Nguyen, G. K. T.; Tam, J. P.; Lescar, J.; Wu, B. Engineering a Catalytically Efficient Recombinant Protein Ligase. J Am Chem Soc 2017, 139 (15), 5351-5358. DOI: 10.1021/jacs.6b12637
  2. [2] (Application, Live cell labelling) Harmand, T. J.; Pishesha, N.; Rehm, F. B. H.; Ma, W.; Pinney, W. B.; Xie, Y. J.; Ploegh, H. L. Asparaginyl Ligase-Catalyzed One-Step Cell Surface Modification of Red Blood Cells. ACS Chemical Biology 2021, 16 (7), 1201-1207. DOI: 10.1021/acschembio.1c00216
  3. [3] (Application, Substrate specificity extension) Rehm, F. B. H.; Tyler, T. J.; Yap, K.; de Veer, S. J.; Craik, D. J.; Durek, T. Enzymatic C-Terminal Protein Engineering with Amines. J Am Chem Soc 2021, 143 (46), 19498-19504. DOI: 10.1021/jacs.1c08976
  4. [4] (Application, peptide cyclization inside cells) Tang, T. M. S.; Mason, J. M. Intracellular Application of an Asparaginyl Endopeptidase for Producing Recombinant Head-to-Tail Cyclic Proteins. JACS Au 2023, 3 (12), 3290-3296. DOI: 10.1021/jacsau.3c00591
  5. [5] (Application, Cyclic peptide display library) Wan, X.-C.; Zhang, Y.-N.; Zhang, H.; Chen, Y.; Cui, Z.-H.; Zhu, W.-J.; Fang, G.-M. Asparaginyl Endopeptidase-Mediated Peptide Cyclization for Phage Display. Organic Letters 2024, 26 (13), 2601-2605. DOI: 10.1021/acs.orglett.4c00602
  6. [6] (Application, Substrate specificity extension) Rehm, F. B. H.; Tyler, T. J.; Zhou, Y.; Huang, Y.-H.; Wang, C. K.; Lawrence, N.; Craik, D. J.; Durek, T. Repurposing a plant peptide cyclase for targeted lysine acylation. Nature Chemistry 2024. DOI: 10.1038/s41557-024-01520-1