TY - JOUR
T1 - Sortase A as a tool for high-yield histatin cyclization
AU - Bolscher, J.G.M.
AU - Oudhoff, M.J.
AU - Nazmi, K.
AU - Antos, J.M.
AU - Guimaraes, C.P.
AU - Spooner, E.
AU - Haney, E.F.
AU - Garcia Vallejo, J.J.
AU - Vogel, H.J.
AU - van 't Hof, W.
AU - Ploegh, H.L.
AU - Veerman, E.C.I.
PY - 2011
Y1 - 2011
N2 - Cyclic peptides are highly valued tools in biomedical research. In many cases, they show higher receptor affinity, enhanced biological activity, and improved serum stability. Technical difficulties in producing cyclic peptides, especially larger ones, in appreciable yields have precluded a prolific use in biomedical research. Here, we describe a novel and efficient cyclization method that uses the peptidyl-transferase activity of the Staphylococcus aureus enzyme sortase A to cyclize linear synthetic precursor peptides. As a model, we used histatin 1, a 38-mer salivary peptide with motogenic activity. Chemical cyclization of histatin 1 resulted in ≤3% yields, whereas sortase-mediated cyclization provided a yield of >90%. The sortase-cyclized peptide displayed a maximum wound closure activity at 10 nM, whereas the linear peptide displayed maximal activity at 10 μM. Circular dichroism and NMR spectroscopic analysis of the linear and cyclic peptide in solution showed no evidence for conformational changes, suggesting that structural differences due to cyclization only became manifest when these peptides were located in the binding domain of the receptor. The sortase-based cyclization technology provides a general method for easy and efficient manufacturing of large cyclic peptides.—Bolscher, J. G. M., Oudhoff, M. J., Nazmi, K., Antos, J. M., Guimaraes, C. P., Spooner, E., Haney, E. F., GarciaVallejo, J. J., Vogel, H. J., van't Hof, W., Ploegh, H. L., Veerman. E. C. I. Sortase A as a tool for high-yield histatin cyclization.
AB - Cyclic peptides are highly valued tools in biomedical research. In many cases, they show higher receptor affinity, enhanced biological activity, and improved serum stability. Technical difficulties in producing cyclic peptides, especially larger ones, in appreciable yields have precluded a prolific use in biomedical research. Here, we describe a novel and efficient cyclization method that uses the peptidyl-transferase activity of the Staphylococcus aureus enzyme sortase A to cyclize linear synthetic precursor peptides. As a model, we used histatin 1, a 38-mer salivary peptide with motogenic activity. Chemical cyclization of histatin 1 resulted in ≤3% yields, whereas sortase-mediated cyclization provided a yield of >90%. The sortase-cyclized peptide displayed a maximum wound closure activity at 10 nM, whereas the linear peptide displayed maximal activity at 10 μM. Circular dichroism and NMR spectroscopic analysis of the linear and cyclic peptide in solution showed no evidence for conformational changes, suggesting that structural differences due to cyclization only became manifest when these peptides were located in the binding domain of the receptor. The sortase-based cyclization technology provides a general method for easy and efficient manufacturing of large cyclic peptides.—Bolscher, J. G. M., Oudhoff, M. J., Nazmi, K., Antos, J. M., Guimaraes, C. P., Spooner, E., Haney, E. F., GarciaVallejo, J. J., Vogel, H. J., van't Hof, W., Ploegh, H. L., Veerman. E. C. I. Sortase A as a tool for high-yield histatin cyclization.
U2 - https://doi.org/10.1096/fj.11-182212
DO - https://doi.org/10.1096/fj.11-182212
M3 - Article
C2 - 21525488
SN - 0892-6638
VL - 25
SP - 2650
EP - 2658
JO - The FASEB Journal
JF - The FASEB Journal
IS - 8
ER -