Three apparently different glucosephosphate isomerases are found in the developing seeds of maize (Zea mays L.). Glu umn chromatography of developing endosperm preparations. Activity under the Ia peak can be enhanced with a concomitant attenuation of activity under the I peak by certain purification procedures. I and II are, however, quite distinct noninterconvertible enzymes that may have different physiological functions. A third enzyme, GPI IIl, is detectable only in the embryo. This enzyme elutes from a DEAE-cellulose column in the same fractions as does GPl lI from the endosperm but is distinguishable from II on the basis of electrophoretic mobility and the stimulatory effect of ATP in the reaction mixtLire.These results contrast with those from pea and green gram (13,16), where highly purified enzymes have been prepared, but no heterogeneity has been reported. GPI from yeast has been crystallized (7). Recently, Nakagawa and Noltmann (8) fractionated the crystallized enzyme into three isozymic forms by elution with a very shallow phosphate gradient from a DEAE-cellulose column. These are stable protein species that are not interconvertible under physiological conditions (9). In contrast to this, Pon et al. (11) have shown that multiple forms of rabbit muscle GPI separated by column chromatography do not remain as single species during rechromatography under the same conditions, although the addition of dithiothreitol prevents previously separated single peaks from separating further. These forms of rabbit muscle isomerases are apparently different forms of the same enzyme protein and can be designated as "pseudoisozvmes." We demonstrate here that both situations are found in developing maize seeds. There are multiple forms that are separable by column chromatography and distinguishable by various tests. Additionally, GPI I can generate a number of electrophoretically separable pseudoisozymes under certain preparative regimens.Since we are interested in the regulation of starch synthesis in maize, glucosephosphate isomerase activity in the maize seed has been investigated. The results reveal that three peaks of activity (I, Ia, II) can be detected after DEAE5-cellulose col-