Modulation of protease activity to enhance the recovery of recombinant nucleocapsid protein of Nipah virus

Structural characterization by transmission electron microscopy and immunoreactivity of recombinant Hendra virus nucleocapsid protein expressed and purified from Escherichia coli

“…This observation is also a potential indicator that the accessibility of the C-terminal tail region increases the susceptibility of the domain to proteolysis and likelihood of intracellular proteolysis when over-expressed in E. coli or other heterologous expression systems. These observations have been made for preparations of HeV N FL protein and NiV N FL expressed in E. coli and S. cerevisiae [34,35,44,45]. Utilizing bioinformatics proteolysis prediction tools, it has been demonstrated that the proteolytic degradation of E. coli expressed NiV N could be reduced by the choice and concentration of specific serine protease inhibitors [35].…”

“…There are many reports on the expression of full-length or truncated constructs of NiV N [23,31,[34][35][36], however, there have only been limited studies on the production of recombinant, full-length HeV N [34,36]. In this report, we describe the expression and purification of soluble full-length HeV N (HeV N FL ) and a C-terminal truncated core domain (HeV N CORE ) lacking the IDR using a bacterial expression systems.…”

“…These observations have been made for preparations of HeV N FL protein and NiV N FL expressed in E. coli and S. cerevisiae [34,35,44,45]. Utilizing bioinformatics proteolysis prediction tools, it has been demonstrated that the proteolytic degradation of E. coli expressed NiV N could be reduced by the choice and concentration of specific serine protease inhibitors [35]. Our bacterial expression studies reveal a high level of proteolysis in the E. coli preparations of HeV N FL , despite the addition of an extensive cocktail of general and serine specific protease inhibitors to the cellular lysis buffer.…”

Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system

“…Like other members of the Paramyxoviridae family, NiV possesses a non‐segmented, negative sense, single‐stranded RNA genome of about 18.2 kb nucleotides . The viral replication and assembly process are located respectively in the cytoplasm and plasma membrane of its host cell, and virions are budded from the cell membrane .…”

“…2,11 Like other members of the Paramyxoviridae family, NiV possesses a non-segmented, negative sense, single-stranded RNA genome of about 18.2 kb nucleotides. 12,13 The viral replication and assembly process are located respectively in the cytoplasm and plasma membrane of its host cell, and virions are budded from the cell membrane. 14 NiV has six transcriptional units in its genome encoding six major proteins: nucleocapsid (N), phospho (P), matrix (M), fusion (F), glyco (G), and large (L).…”