An Improved Method for the Isolation of Carboxydismutase. Probable Identity with Fraction I Protein and the Protein Moiety of Protochlorophyll Holochrome<sup>*</sup>

Trown, P. W.

doi:10.1021/bi00881a018

Cited by 136 publications

(52 citation statements)

References 36 publications

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“…The production of these enzymes also showed differing temperature optima in leaves of a temperature-sensitive chlorophyll mutant of alfalfa (6). Present evidence shows that fraction I protein is probably crude ribulose-l,5-diP carboxylase (15,16). Fraction I protein may account for up to 50% of the soluble protein of green leaves (3) and is located in the chloroplast (11).…”

Section: Resultsmentioning

confidence: 68%

Influence of Age and Illumination on Distribution of Several Calvin Cycle Enzymes in Greening Barley Leaves

Obendorf¹,

Huffaker²

1970

Plant Physiol.

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Activities of phosphoriboisomerase, phosphoribulokinase, and ribulose 1,5-diphosphate carboxylase, protein content, and chlorophyll accumulation in dark-grown barley seedlings were measured before and after illumination. Enzymatic activities, levels of soluble protein, and accumulation (upon illumination) of chlorophyll in leaves declined from tips toward the base. In response to increasing time of illumination, chlorophyll accumulation and activities of phosphoribulokinase and ribulose 1 ,5-diphosphate carboxylase (enzymes located in chloroplasts) increased most in tip portions whereas activity of phosphoriboisomerase and levels of soluble protein (constituents not confined to chloroplasts) increased similarly in all sections of the leaf. Maximum activity of phosphoribulokinase and maximum accumulation of chlorophyll shifted toward median portions of the leaf blade with increased age of seedling before illumination. Maximum activity of ribulose 1 ,5-diphosphate carboxylase and maximum level of soluble protein occurred in all leaf sections when the seedlings were 7 days of age before illumination.Upon the illumination of dark-grown seedlings, chlorophyll accumulates, protein is synthesized, chloroplasts develop an ordered lamellar form, the activity of photosynthetic enzymes increases, and net incorporation of CO2 begins. Some information is available on the development of several of these and other components with position in the leaf and age of the leaf. Rhodes and Yemm (13) reported that chloroplasts were larger and developed faster in the older cells of barley leaf tips than in younger cells at the base. Protochlorophyll levels in etiolated barley leaves declined from the tip toward the base (17). A decline in the tip with increasing age, however, shifted the maximal concentration toward the median sections. Williams and Rijven (19) noted marked differences in the timing of maximum protein and RNA levels for different parts of developing wheat leaves. Information is lacking on the development of photosynthetic enzymes in relation to position in the leaf and the development of other constituents. This paper reports the influence of illumination and leaf age and section on the activities of phosphoriboisomerase, phosphoribulokinase, and ribulose-1,5-diP carboxylase, and on soluble protein and chlorophyll.'Present address:

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Section: Resultsmentioning

confidence: 68%

Influence of Age and Illumination on Distribution of Several Calvin Cycle Enzymes in Greening Barley Leaves

Obendorf¹,

Huffaker²

1970

Plant Physiol.

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“…We have considered the possibility that Group I represents solely the large subunit of RuDP-carboxylase, which has a mol wt of 55,000 (11). However, carboxylase has been shown to be removed from the chloroplast membranes of spinach by washing with water (35) and is routinely prepared from homogenates obtained with the aid of a French pressure cell without further extraction of the membranes (11,36). Furthermore, we have performed several experiments which bear on the question of the nature of the Group I polypeptides.…”

Section: Discussionmentioning

confidence: 99%

SYNTHESIS OF CHLOROPLAST MEMBRANE POLYPEPTIDES DURING SYNCHRONOUS GROWTH OF CHLAMYDOMONAS REINHARDTII

Beck

Levine

1974

The Journal of Cell Biology

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The synthesis of the major chloroplast membrane polypeptides has been studied during synchronous growth of Chlamydomonas reinhardtii. Under these conditions, chlorophyll is synthesized during the latter part of the light period and cell division takes place during the dark period. The profile of the chloroplast membrane polypeptides of C. reinhardtii has been well characterized and shown to contain two major classes by size (Hoober, J. 1970. J. Biol. Chem. 245:4327). Polypeptides of group I have a mol wt range of 50,000-55,000 daltons. The second region consists of at least three polypeptide groups, IIa, lib, and IIc, having mol wt of 40,000, 31,000, and 27,000 daltons, respectively. The synthesis of these polypeptides has been measured using a doublelabeling technique and a computer-aided statistical analysis. The rate of labeling of group I polypeptides is highest during the early light period and decreases after 6 h of growth. Group IIa is labeled from the beginning of the light period, but little synthesis of lib occurs before 3 h, and significant amounts of label are not found in IIc before 5 h of growth. After approximately 8 h of light, groups lib and IIc are synthesized at rates significantly greater than those of the other membrane polypeptides. The synthesis of the major polypeptide groups ceases in the dark. We conclude that the biosynthesis of the chloroplast membranes is a sequential or stepwise process.Much information has recently become available concerning the physical properties of biological membranes. Less, however, is known about the processes of synthesis and integration of components required for the production of functional membranes, especially in eukaryotic cells. First, it is not known whether there exists an obligatory coupling of the synthesis of one or more of the proteins and lipids involved in the formation of membranes under conditions of normal cell growth, and, second, it is not known whether the association of specific lipid and protein components is required for their integration into the membrane. With regard to the first question, experiments with bacterial mutants have indicated that the synthesis of membrane lipids and proteins is closely coordinated (1-4). On the other hand, the turnover of lipids and proteins of rat liver cell endoplasmic reticulum appears to occur at different rates (5, 6), and studies with the y-I mutant of Chlamydomonas reinhardtii have shown that the ratios of various chloroplast membrane components vary during the greening process (7 9). These results sug-

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“…Below a limiting load, the response of the spectrophotometer in terms of weight of chart paper under each peak was proportional to the amount of RNA or protein. The weight of chart paper was related to the amounts of RNA or fraction I protein using published extinction coefficients: that for rRNA was taken to be E2,n'm = 223 (23) and that for fraction I protein to be E211,m =14.1 (24).…”

Section: Methodsmentioning

confidence: 99%

“…4). Fraction I protein, which is identical to the enzyme ribulose 1, 5-diphosphate carboxylase (24), is probably made on the chloroplast ribosomes by the alga Euglena (20).…”

Section: Methodsmentioning

confidence: 99%

Developmental Changes in Ribosomal Ribonucleic Acid and Fraction I Protein in Wheat Leaves

Patterson

Smillie

1971

Plant Physiol.

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In light-grown wheat (Triticumn aestivum L.) seedlings, the amount of chloroplast and cytoplasmic ribosomal RNA increased to a maximum in the first leaf near the end of its growth and declined by about 60% in the following 3 days. While total ribosomal RNA was declining, labeled uracil was still incorporated into cytoplasmic ribosomal RNA, but the rate of incorporation into chloroplast ribosomal RNA fell by more than 80%, as did the incorporation of labeled leucine into fraction I protein. Either there is greater replacement of cytoplasmic ribosomal RNA than chloroplast ribosomal RNA in mature leaves, or chloroplasts are able to repress the incorporation of exogenous precursor when there is no net synthesis of RNA.In leaves there are two major systems of protein synthesis (2, 18), one using the ribosomes of the cytoplasm, and the other using the smaller ribosomes of the chloroplast (4,19,20). Although it is difficult to estimate relative amounts of the two classes of ribosomes directly, they each contain characteristic species of RNA which can be separated and estimated using polyacrylamide gel electrophoresis (6,8). The larger subunit of chloroplast rRNA in particular is somewhat unstable during isolation (6). Therefore, in order to study the net changes in rRNA at different stages of the development of wheat leaves, we have measured the amounts of the smaller subunits of chloroplast and cytoplasmic rRNA. We have compared the changes in rRNA levels with changes in the level of fraction I protein, which is probably made on the chloroplast ribosomes (20). In addition, the ability of wheat leaves to incorporate exogenous precursor into chloroplast rRNA, cytoplasmic rRNA, and fraction I protein has been measured at different stages of development. MATERIALS AND METHODSGrowth of Wheat. Wheat (Triticum aestivum L., var. Timgalen) was grown in vermiculite with a 24 hr cycle of 12-hr light and dark, at 22 C and 16 C, respectively. Light (about 9,000 lux) was supplied by equal numbers of Philips MCFER warm white and Sylvania Grolux fluorescent tubes. Sowing and harvesting was midway through the light period, and the nutrient solution used was that of Hoagland and Arnon (5).Incorporation of Radioisotopes. Leaves were dissected from the coleoptile and cut at the ligule, surface sterilized with 0.1% cetyltrimethylammonium bromide, thoroughly rinsed with sterile distilled water, and cut into 3-mm sections. Tissue (0.5 g) was incubated with 1 ml of precursor solution for 6 hr with intermittent shaking in light at 22 C. 5-3H-Uracil (240 ,uc/ml; 1 c/mmole) was used as a precursor of RNA, and 'H-leucine, generally labeled (100 ,uc/ml; 50 mc/mmole), was used as a precursor of protein. Incubation was begun 3 hr before the middle of the light period.Extraction of RNA. RNA was prepared using Method B of Loening and Ingle (8) and purified by the method of Ralph and Bellamy (17). Total RNA and DNA were estimated by the method of Smillie and Krotkov (21).Extraction of Fraction I Protein. Leaves were frozen in liquid nitrogen a...

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An Improved Method for the Isolation of Carboxydismutase. Probable Identity with Fraction I Protein and the Protein Moiety of Protochlorophyll Holochrome^*

Abstract: of protoehlorophyllholochroma. •me purified on~;me ahowe~ no s1f,ylificant i phosphor1bo1nomm•a.sa or phosphor1bu.lo~lna.$e act1 Vities; thes

Cited by 136 publications

References 36 publications

Influence of Age and Illumination on Distribution of Several Calvin Cycle Enzymes in Greening Barley Leaves

Influence of Age and Illumination on Distribution of Several Calvin Cycle Enzymes in Greening Barley Leaves

SYNTHESIS OF CHLOROPLAST MEMBRANE POLYPEPTIDES DURING SYNCHRONOUS GROWTH OF CHLAMYDOMONAS REINHARDTII

Developmental Changes in Ribosomal Ribonucleic Acid and Fraction I Protein in Wheat Leaves

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An Improved Method for the Isolation of Carboxydismutase. Probable Identity with Fraction I Protein and the Protein Moiety of Protochlorophyll Holochrome*

Abstract: of protoehlorophyllholochroma. •me purified on~;me ahowe~ no s1f,ylificant i phosphor1bo1nomm•a.sa or phosphor1bu.lo~lna.$e act1 Vities; thes

Cited by 136 publications

References 36 publications

Influence of Age and Illumination on Distribution of Several Calvin Cycle Enzymes in Greening Barley Leaves

Influence of Age and Illumination on Distribution of Several Calvin Cycle Enzymes in Greening Barley Leaves

SYNTHESIS OF CHLOROPLAST MEMBRANE POLYPEPTIDES DURING SYNCHRONOUS GROWTH OF CHLAMYDOMONAS REINHARDTII

Developmental Changes in Ribosomal Ribonucleic Acid and Fraction I Protein in Wheat Leaves

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An Improved Method for the Isolation of Carboxydismutase. Probable Identity with Fraction I Protein and the Protein Moiety of Protochlorophyll Holochrome^*