title: Re-examining the oligomerization state of macrophage migration inhibitory factor (MIF) in solution.
abstract: The state of oligomerization of macrophage migration inhibitory factor (MIF, also known as glycosylation inhibiting factor, GIF) in solution has been variously reported as monomer, dimer, trimer, or mixtures of all three. Several crystal structures show MIF to be a trimer. Sedimentation velocity shows a recombinant human MIF sample is quite homogeneous, with 98% as a species with s(20,w)=3.07 S and D(20,w)=8.29 x 10(-7) cm(2)/s. Using the partial specific volume calculated from the amino acid composition these values imply a mass of 33.56 kDa, well above that of dimer, but also 9% below the trimer mass of 37.035 kDa. Sedimentation equilibrium data at loading concentrations from 0.01 to 1 mg/ml show unequivocally that the self-association is extremely tight. However, the apparent mass is 33.53 kDa [95% confidence 33.25-33.82], again 9% below that expected for 100% trimer. To examine the possibility this protein has an unusual partial specific volume, sedimentation equilibrium was also done in H(2)O/D(2)O mixtures, giving 0.765+/-0.017 ml/g rather than the calculated 0.735 ml/g. With this revised partial specific volume, the equilibrium and velocity data each give M=37.9+/-2.8 kDa, fully consistent with a strongly-associated trimeric quaternary structure.
authors.affiliation: Alliance Protein Laboratories, 3957 Corte Cancion, Thousand Oaks, CA 91360, USA. jphilo@mailway.com
authors.forename: John S
authors.initials: JS
authors.lastname: Philo
authors.forename: Tzung-Horng
authors.initials: TH
authors.lastname: Yang
authors.forename: Michael
authors.initials: M
authors.lastname: LaBarre
mesh_terms: D018360:Crystallography, X-Ray; D006801:Humans; D008263:Macrophage Migration-Inhibitory Factors / Q000737:chemistry*; D013058:Mass Spectrometry; D008970:Molecular Weight; D011108:Polymers / Q000737:chemistry; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry; D012996:Solutions; D014461:Ultracentrifugation
chemical_list: D008263:Macrophage Migration-Inhibitory Factors; D011108:Polymers; D011994:Recombinant Proteins; D012996:Solutions