Expression of the superantigen Mycoplasma arthritidis mitogen in Escherichia coli and characterization of the recombinant protein.
Mycoplasma arthritidis mitogen (MAM), is a soluble protein with classical superantigenic properties and is produced by an organism that causes an acute and chronic proliferative arthritis. Unfortunately, the process of obtaining purified MAM from M. arthritidis culture supernatants is extremely time-consuming and costly, and very little material is recovered. Thus, our laboratory has expressed MAM in Escherichia coli by using a protein fusion expression system. The construction and expression of recombinant MAM (rMAM), as well as a comparison of the biological properties of rMAM to those of native MAM, are discussed. Briefly, conversion of the three UGA codons to UGG codons was required to obtain full-length expression and mitogenic activity of rMAM. Antisera to native MAM recognized both rMAM and the fusion protein. The T-cell receptor Vbeta and major histocompatibility complex class II receptor usages by rMAM and the fusion protein were identical to that of native MAM. In addition, the ability to induce suppression and form the superantigen bridge could also be demonstrated with rMAM. Importantly, dose-response experiments indicated that homogeneous native MAM and rMAM were of equal potency. Thus, MAM has been successfully expressed in E. coli, thereby creating a viable alternative to native MAM.
Knudtson
K L
KL
Department of Internal Medicine, University of Utah School of Medicine, Salt Lake City 84132, USA.
Manohar
M
M
Joyner
D E
DE
Ahmed
E A
EA
Cole
B C
BC
D000941:Antigens; D000942:Antigens, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D008934:Mitogens / Q000235:genetics*; D011506:Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics; D018089:Superantigens / Q000235:genetics*
D000941:Antigens; D000942:Antigens, Bacterial; D008934:Mitogens; C052996:Mycoplasma arthritidis mitogen; D011506:Proteins; D011993:Recombinant Fusion Proteins; D018089:Superantigens