Total synthesis of a functional SUMO-1 peptide conjugate.
Nathalie Ollivier, Emmanuelle Boll, Hervé Drobecq, Laurent Raibaut, Jérôme
Vicogne* and Oleg Melnyk.
UMR CNRS 8161 CNRS, Université de Lille, Institut Pasteur de Lille, 1 rue du Pr
Calmette, 59021 Lille Cedex, France.
The large diversity of chemical methods available now for producing modified
peptides in high purity is increasingly used for the total or semisynthesis of modified
proteins which cannot be easily produced using biological methods. Proteins
featuring well-defined post-translational modifications at specific sites such as
ubiquitylation belong to this family. The total synthesis of small ubiquitin-like modifier
(SUMO) proteins and SUMO conjugates is challenging and is addressed in this work.
Indeed, we describe for the first time the total synthesis of a folded, functional and
reversible 12.1 kD SUMO-1 conjugate (Figure),1-2 which was assembled using a onepot N-to-C three segments assembly process relying on sequential NCL and SEA
ligation reactions.3-6
The assembly of SUMO-1 proteins required to push the limits of current
technology by addressing the synthesis of long (~ 50 AA) SEA peptides and peptide
thioesters. This was achieved by developing a novel SEA PEG-based solid support
which is compatible with standard Fmoc-SPPS.
The tertiary structure and functionality of SUMO-1 modifier within the
conjugate as well as its reversible nature was demonstrated by the total and specific
cleavage of the conjugate with a recombinant SUMO-1 deconjugating enzyme.
MALDI-TOF
RP-HPLC
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E. Boll, H. Drobecq, N. Ollivier, L. Raibaut, R. Desmet, J. Vicogne and O.
Melnyk, Chem. Sci., 2014, 5, 2017-2022.
E. Boll, H. Drobecq, N. Ollivier, A. Blanpain, L. Raibaut, R. Desmet, J.
Vicogne and O. Melnyk, Nat. Protocols, 2015, 10, 269-292.
N. Ollivier, J. Dheur, R. Mhidia, A. Blanpain, O. Melnyk, Org. Lett., 2010, 12,
5238 (2010)
N. Ollivier, J. Vicogne, A. Vallin, H. Drobecq, R. Desmet, O. El-Mahdi, B.
Leclercq, G. Goormachtigh, V. Fafeur and O. Melnyk, Angew. Chem., Int. Ed.,
2012, 51, 209-213.
L. Raibaut, N. Ollivier, O. Melnyk, Chem. Soc. Rev., 2012, 41, 7001.
L. Raibaut, J. Vicogne, B. Leclercq, H. Drobecq, R. Desmet, O. Melnyk,
Bioorg. Med. Chem., 2013, 21, 3486.