PURIFICATION AND DIFFERENTIAL EXPRESSION OF ENOLASE FROM MAIZE

Enolase was purified from maize (Zea mays L. inbred B73) seeds to a 55 and 56 kDa protein doubler based upon sodium dodecyl sulfate-polyacry lamide gel electrophoresis. Purification included ammonium sulfate-pre cipitation, gel filtration, Mono Q, and Phenyl Superose chromatography . Two-dimensional gels further resolved the 56 kDa protein into three isoelectric forms. Polyclonal antibodies raised against the purified p roteins, were found to bind specifically to both the 55 and 56 kDa pro teins during purification. These antibodies did not bind protein in ce ll extracts of DF261 (an enolase deficient mutant of E. coli), but rec ognized a 56 kDa protein when the strain was complemented with maize e nolase (pZM245). Maize enolase antibodies recognized a similar 56 kDa protein in several plant species. A comparison of maize shoot and root extracts indicated that the 55 kDa form of enolase was more abundant in roots. Enolase protein levels remained unchanged in maize roots aft er 24 h of anaerobiosis, even though the specific activity of enolase increased to twice its initial levels. A plastid form of enolase in ma ize could not be found as either enolase activity or protein (with imm unoblots).