TY - JOUR
T1 - Addition of magnesium chloride to enhance mono-dispersity of a coiled-coil recombinant mouse macrophage protein
AU - Pahuja, Parveen
AU - Srinivasan, Alagiri
AU - Puri, Munish
PY - 2014/4
Y1 - 2014/4
N2 - X-ray crystallography for the determination of three-dimensional structures of protein macromolecules represents an important tool in function assignment of uncharacterized proteins. However, crystallisation is often difficult to achieve. A protein sample fully characterized in terms of dispersity may increase the likelihood of successful crystallisation by improving the predictability of the crystallisation process. To maximize the probability of crystallisation of a novel mouse macrophage protein (rMMP), target molecule was characterized and refined to improve monodispersity. Addition of MgCl 2 at low concentrations resolves the rMMP into a monodisperse solution, and finally successful crystallization of rMMP was achieved. The effect of MgCl2 was studied using gel filtration chromatography and dynamic light scattering.
AB - X-ray crystallography for the determination of three-dimensional structures of protein macromolecules represents an important tool in function assignment of uncharacterized proteins. However, crystallisation is often difficult to achieve. A protein sample fully characterized in terms of dispersity may increase the likelihood of successful crystallisation by improving the predictability of the crystallisation process. To maximize the probability of crystallisation of a novel mouse macrophage protein (rMMP), target molecule was characterized and refined to improve monodispersity. Addition of MgCl 2 at low concentrations resolves the rMMP into a monodisperse solution, and finally successful crystallization of rMMP was achieved. The effect of MgCl2 was studied using gel filtration chromatography and dynamic light scattering.
KW - Crystallisation
KW - Dispersity
KW - Dynamic light scattering
KW - Gel filtration chromatography
UR - http://www.scopus.com/inward/record.url?scp=84896543315&partnerID=8YFLogxK
U2 - 10.1007/s11010-013-1934-x
DO - 10.1007/s11010-013-1934-x
M3 - Article
VL - 389
SP - 133
EP - 139
JO - MOLECULAR AND CELLULAR BIOCHEMISTRY
JF - MOLECULAR AND CELLULAR BIOCHEMISTRY
SN - 0300-8177
IS - 1-2
ER -