Identification of Peroxisome Membrane Proteins (PMPs) in Sunflower (Helianthus annuus L.) Cotyledons and Influence of Light on the PMP Developmental Pattern

Abstract

Boundary membranes were recovered from glyoxysomes, transition peroxisomes, and leaf-type peroxisomes purified from cotyledons of sunflower (Helianthus annuus 1.) at three stages of postgerminative growth. After membranes were washed in 100 mM Na2C03 (pH 11.5), integral peroxisome membrane proteins (PMPs) were solubilized in buffered aminocaproic acid/dodecyl maltoside (0.63 ~/1.5%) and analyzed by nondenaturing and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Six prominent nondenatured PMP complexes and 10 prominent SDS-denatured polypeptides were identified in the membranes of the three types of peroxisomes. A nondenatured complex of approximately 140 kD, composed mainly of 24.5-kD polypeptides, decreased temporally, independently of seedling exposure to white, blue, or red light; only far-red light seemed to prevent its decrease. PMP complexes of approximately 120 and 70 kD, in contrast, were present at all stages and changed in polypeptide content. It remains to be determined whether these data reflect changes within in vivo complexes or within complexes formed following/during detergent solubilization. Conversion of glyoxysomes to leaf-type peroxisomes in white or red light after a 2-d dark period was accompanied by the appearance of three SDS-denatured PMPs: 27.5, 28, and 47 kD. The former two became part of the PMPlZO and 70 complexes, as well as part of a new PMP130 complex that also possessed the PMP47. Growth of seedlings in blue or far-red light did not promote the appearance of PMPs 27.5 or 28. Blue light promoted the appearance of PMP47, and far-red light seemed to prevent its appearance. Chlorophyll likely is not the photoreceptor involved in accumulation of PMPs because the PMP composition is distinctly different in seedlings irradiated with red or blue light of comparable fluence rates. Several lines of evidence indicate that the synthesis and acquisition of membrane and all matrix proteins are not coupled. The data provide evidence for a change in PMP composition when sunflower or any other oilseed glyoxysomes are converted to leaf-type peroxisomes and suggest that the change is regulated by both photobiological and temporal mechanisms.Germinated oilseeds are an excellent system in which to study the developmental biogenesis of peroxisomes. Following germination, oil stored in cotyledons are converted mostly

Copper–zinc superoxide dismutase is a constituent enzyme of the matrix of peroxisomes in the cotyledons of oilseed plants

“…This biochemical evidence, together with the results from electron microscopy (Corpas et al, 1994), show that cucumber peroxisomes have their own Mn-SOD. However, in the case of the sunflower organelles (Figs 1 b, 2b), the activities of the marker enzyme indicated some cross-contamination between peroxisomes and mitochondria (Jiang et al, 1994). From comparisons of the densitometric CuZn-SOD staining activities (Figs 1 b, 2b), the peroxisomal CuZn-SOD activities could be attributed to isoforms III and IV, whilst the mitochondrial CuZn-SOD constituted isoforms I and II.…”

“…Organelles were isolated from cotyledons of 2-d dark-grown seedlings of cucumber, cotton or sunflower by differential and linear (cucumber, cotton), or step (sunflower), density-gradient centrifugations in sucrose, as previously described by Corpas, Bunkelmann & Trelease (1994), Chapman & Trelease (1991), and Jiang et al (1994, respectively. The organelle fractions were highly purified as determined by biochemical marker enzyme assays and electron microscopy (Chapman & Trelease, 1991 ;Corpas et al, 1994 ;Jiang et al, 1994).…”

“…The organelle fractions were highly purified as determined by biochemical marker enzyme assays and electron microscopy (Chapman & Trelease, 1991 ;Corpas et al, 1994 ;Jiang et al, 1994).…”

Peroxisomes Are Required for in Vivo Nitric Oxide Accumulation in the Cytosol following Salinity Stress of Arabidopsis Plants

“…Gels were stained with silver according to the modifications described in detail by Jiang et al (1994) for the Heukeshoven and Dernick method (Heukeshoven and Dernick, 1985). For western-blot analysis, proteins were electroblotted to polyvinylidene difluoride membranes with a semidry Trans-Blot cell (Bio-Rad).…”

Plasma Membrane Intrinsic Proteins of Beta vulgaris L

“…Other MIPs have now been identified in plant tissue. These include RD28 (Yamaguchi-Shinozaka et al, 1992), a water-stress protein from Arubidopsis, TRAMP (or pTOM75) (Fray et al, 19941, a group of peroxisomal membrane proteins (Corpas et al, 1994;Jiang et al, 1994), and two families of integral proteins (PIP 1 and 2) derived from the PM of Arubidopsis (Kammerloher et al, 1994). These latter proteins were cloned by immunoselection using a mammalian expression system and were shown to be water channels by osmotic water permeability studies in a Xenopus oocyte system (Kammerloher et al, 1994).…”