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Cis autocatalytic cleavage of glycine‐linked Zika virus NS2B‐NS3 protease constructs
Author(s) -
Hammerstein Franziska,
Lauth Luca M.,
Hammerschmidt Stefan,
Wagner Annika,
Schirmeister Tanja,
Hellmich Ute A.
Publication year - 2019
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13507
Subject(s) - ns3 , protease , ns2 3 protease , serine protease , cleavage (geology) , chemistry , linker , masp1 , virology , biology , biochemistry , enzyme , paleontology , fracture (geology) , computer science , operating system
The flaviviral heterodimeric serine protease NS2B‐NS3, consisting of the NS3 protease domain and the NS2B co‐factor, is essential for ZIKA virus maturation and replication in cells. For in vitro studies a ‘linked’ construct, where a polyglycine linker connects NS2B CF and NS3 pro , is often used. This construct undergoes autocatalytic cleavage. Here, we show that linked ZIKV NS2B CF ‐NS3 pro is cleaved in cis in the NS2B CF exclusively at position R95 and not at the previously proposed alternate cleavage site at residue R29 in the NS3 pro . Cleavage neither affects protease stability nor activity, despite some observed differences in spectroscopic behavior. This minimally modified construct may thus be useful for future structural and functional studies of the flaviviral protease, for example when testing new inhibitors.