Chloroplast Biogenesis: XXIV. Intrachloroplastic Localization of the Biosynthesis and Accumulation of Protoporphyrin IX, Magnesium-Protoporphyrin Monoester, and Longer Wavelength Metalloporphyrins during Greening.

The intraplastidic localization of the endogenous metabolic pools from protoporphyrin to protochlorophyll was determined in Cucumis sativus. The endogenous protoporphyrin, Mg-protoporphyrin monoester + longer wavelength metalloporphyrins, protochlorophyllide and protochlorophyllide ester were membrane-bound. Protoporphyrin was synthesized in the stroma and subsequently became associated with the membranes. The membrane-associated protoporphyrin was then converted into Mg-protoporphyrin monoester + longer wavelength metalloporphyrins by membrane-bound enzymes. Although lysed plastids were capable of converting exogenous delta-aminolevulinic acid to protochlorophyllide, the net synthesis of protochlorophyllide from exogenous delta-aminolevulinic acid was lost upon segregating the lysed plastids into stromal and membrane fractions and then recombining the stromal and membrane fraction prior to incubation. The segregated membrane fraction was still capable of converting protoporphyrin into Mg-protoporphyrin monoester + longer wavelength metalloporphyrins in the presence or absence of the stromal fraction. These results indicated that although the reactions from protoporphyrin to Mg-protoporphyrin monoester and longer wavelength metalloporphyrins could survive a considerable degree of plastid disruption, the reactions from Mg-protoporphyrin monoester and longer wavelength metalloporphyrins to protochlorophyllide were more sensitive to structural disorganization.