Arch Virol (2003) 148: 1301–1316
Biochemical and biophysical analysis of heptad repeat regions
from the fusion protein of Menangle virus,
a newly emergent paramyxovirus
J. Q. Zhu
, C. W.-H. Zhang
, P. Tien
, and G. F. Gao
Department of Molecular Virology, Institute of Microbiology,
Chinese Academy of Sciences, Beijing, China
Oxagen Ltd., Milton Park, Oxfordshire, U.K.
Laboratory of Structural Biology and MOE Laboratory of Protein Sciences,
School of Life Sciences and Bio-Engineering, Tsinghua University, Beijing, China
Nufﬁeld Department of Clinical Medicine, John Radcliffe Hospital,
University of Oxford, Oxford, U.K.
Received January 23, 2003; accepted February 28, 2003
Published online April 28, 2003
Summary. Menangle virus is a novel paramyxovirus isolated in Australia in 1997,
but its classiﬁcation position has not yet been ﬁnally settled. Here by using a
computational program, LearnCoil-VMF, we determined the heptad repeat (HR)
regions (HR1 and HR2) of Menangle virus F protein. Subsequently the HR1
and HR2 peptides were expressed as a single chain (named 2-Helix) connected
by a six amino-acid linker as a GST fusion protein with an E. coli in vitro
expression system. The GST-removed puriﬁed 2-Helix protein could form a stable
trimer in vitro judging by gel-ﬁltration and chemical cross-linking. CD spectra
showed that the 2-Helix protein had a high percentage of α-helix and was very
thermo-stable. Crystals of the 2-Helix protein preparations have been obtained
in many conditions with hanging-drop diffusion method. These results indicated
that Menangle virus has the common features of the fusion protein for other
paramyxoviruses and should adopt a similar fusion mechanism to other members.
As the HR regions of Menangle virus F protein could form stable six-helix
bundle coiled coil structure, they should be used as drug target for the design of
fusion inhibitors, as successfully used for other parmyxoviruses. This is especially
relevant to such a newly emergent virus with zoonotic potentials.
Menangle virus (MenV) is a newly isolated paramyxovirus from stillborn piglets
at a large commercial piggery in New South Wales, Australia in 1997 .