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Biomedical subjects

T W Goodwin

Publications and source records attributed to T W Goodwin.

At least 91 records · Page 5Linked to original sources

Incorporation of 2-[14C]mevalonic acid into phytoene by isolated chloroplasts.

1. Chloroplasts prepared by the non-aqueous technique will, after fragmentation by ultrasonic treatment, incorporate [2-(14)C]mevalonic acid into phytoene, the first C(40) compound formed in the biosynthetic sequence to coloured carotenoids. 2. With suspensions containing 3.5mg. of chlorophyll, the optimum amounts of cofactor required were ATP (10mumoles), magnesium chloride (20mumoles) and glutathione (20mumoles); neither NAD(+) nor NADP(+) was required. 3. Very small amounts of squalene are also formed and synthesis is stimulated by addition of NADH or NADPH. Phytoene synthesis was not affected by the presence of these cofactors and no lycopersene (the C(40) homologue of squalene) was detected. 4. The phytol side chain of chlorophyll is also labelled under these conditions. 5. Preparations of developing chloroplasts are more active than preparations of mature chloroplasts.

Adenosine Triphosphate↗

Observations on the biosynthesis of thiamine in yeast.

1. Methods are described for the isolation of radioactively pure thiamine from yeast and its degradation on a small scale to its cyclic components. 2. A degradation of the pyrimidine ring and a thin-layer method for the separation of thiamine, its derivatives and pyrimidine and thiazole residues are described. 3. [(14)C]Formate is more effectively incorporated into the pyrimidine residue than into the thiazole residue, whereas the reverse is true with l-[Me-(14)C]methionine. 4. Experiments with [Me-(14)C,(35)S]methionine demonstrate that methionine provides an intact unit for the biosynthesis of the thiazole ring. 5. [6-(14)C]Orotic acid is insignificantly incorporated into the pyrimidine residue of thiamine. 6. Experiments with [1-(14)C]- and [2-(14)C]-acetate indicate that it is incorporated as a unit into the thiazole residue, but that only C-2 is incorporated into the pyrimidine residue. 7. l-[U-(14)C]Alanine is also effectively incorporated into the thiazole residue. 8. These results are discussed in relation to possible pathways of biosynthesis of the two ring components of the thiamine molecule.

Acetates↗

Some observations on the biosynthesis of the plant sulpholipid by Euglena gracilis.

1. dl-Cysteine decreases the uptake of (35)SO(4) (2-) by Euglena gracilis but does not decrease the relative incorporation of the isotope into sulpholipid; cysteic acid, on the other hand, does not affect the uptake of (35)SO(4) (2-) but does dilute out its incorporation into the sulpholipid. 2. Both l-[(35)S]cysteic acid and dl-+meso-[3-(14)C]cysteic acid appear almost exclusively in 6-sulphoquinovose. 3. Molybdate inhibits the incorporation of (35)SO(4) (2-) into sulpholipid but not its uptake into the cells; this suggests that adenosine 3'-phosphate 5'-sulphatophosphate may be concerned with the biosynthesis of sulpholipid, and it was shown to be formed by chloroplast fragments. 4. An outline scheme for sulpholipid biosynthesis based on these observations is discussed.

Chloroplasts↗

Incorporation of [14C] carbon dioxide and [2-14C] mevalonic acid into terpenoids of higher plants during chloroplast development.

1. The incorporation of (14)CO(2) and dl-[2-(14)C]mevalonic acid into various terpenoids in developing chloroplasts in a number of seedlings has been studied. 2. beta-Carotene and phytol (from chlorophyll) tend to be heavily labelled from (14)CO(2), whereas sterols and beta-amyrin are only slightly labelled; with dl-[2-(14)C]mevalonic acid the situation is reversed. 3. The incorporation of (14)CO(2) into terpenoids is dependent on the stage of chloroplast development, whereas that of mevalonic acid is independent of chloroplast development. 4. The uptake of (14)CO(2) into beta-carotene and phytol in mature chloroplasts is very low in monocotyledons but somewhat greater in dicotyledons. 5. The results are discussed in relation to the view that terpenoid biosynthesis in developing chloroplasts is regulated by a combination of enzyme segregation and specific membrane permeability.

Carbon Dioxide↗

Intracellular localization of mevalonate-activating enzymes in plant cells.

Mevalonate-activating enzymes are shown to be present in the chloroplasts of French-bean leaves. The chloroplast membrane is impermeable to mevalonic acid. Mevalonate-activating enzymes also appear to be found outside the chloroplast. These results support the view that terpenoid biosynthesis in the plant cell is controlled by a combination of enzyme segregation and specific membrane permeability.

Autoradiography↗

The stereospecific biosynthesis of plant sterols and alpha- and beta-amyrin.

1. A preparation of pea seedlings has been obtained that will incorporate [2-(14)C]mevalonate into squalene, alpha- and beta-amyrin and the phytosterols. 2. The (14)C/(3)H ratio in alpha- and beta-amyrin biosynthesized in the presence of [2-(14)C,4R-(3)H]-mevalonate is the same as in the starting material and in squalene; this gives experimental support to the mechanism for the cyclization of squalene proposed by Ruzicka for the formation of these pentacyclic triterpenoids. 3. The (14)C/(3)H ratio for beta-sitosterol was 5:3, the same as that in cholesterol in liver. 4. As the absence of (3)H from C-3 in beta-sitosterol was demonstrated (3)H must be present in the side chain and thus the H at C-24 is not lost during alkylation of the side chain; it probably migrates to C-25.

Carbon Isotopes↗

The biosynthesis of sterols in higher plants.

1. [2-(14)C]Mevalonate was incorporated into squalene and the major phytosterols of pea and maize leaves; it was also incorporated into compounds belonging to the 4,4-dimethyl and 4alpha-methyl steroid groups and which may be possible phytosterol intermediates. 2. l-[Me-(14)C]Methionine was incorporated into the major sterols and also into the 4,4-dimethyl and 4alpha-methyl steroid groups. No radioactivity was detected in squalene. 3. Under anaerobic conditions incorporation of [2-(14)C]-mevalonate into the non-saponifiable lipid of pea leaves was drastically decreased but radioactive squalene was accumulated. 4. Cycloartenol, 24-methylenecycloartanol, 24-methylenelophenol, 24-ethylidenelophenol, fucosterol, beta-sitosterol, stigmasterol and campesterol have been identified by gas-liquid chromatography in pea leaves. 5. The significance of these results in connexion with phytosterol biosynthesis and the introduction of the alkyl group at C-24 into phytosterols is discussed.

Carbon Isotopes↗