Improved resolution of hydrophobic penicillin-binding proteins and their covalently linked complexes on modified C18 reversed phase column

Hydrophobic proteins have proven to be difficult to separate chromatographi cally. The use of a C18 reversed phase column with embedded polar groups sh owed improved separation and recovery of the hydrophobic, membrane associat ed, penicillin-binding proteins of Bacillus subtilis and their covalently l inked complexes. The covalently linked complexes were generated by a time c ourse reaction of the non-covalently associated complexes in the intact cel l with the paired group specific reagent, cyanogen (C2N2).