Biomedical subjects

S M Theg

Publications and source records attributed to S M Theg.

21 records · Page 2Linked to original sources

PubMed1988

Isolation of a cDNA clone for the gamma subunit of the chloroplast ATP synthase of Chlamydomonas reinhardtii: import and cleavage of the precursor protein.

A cDNA library from Chlamydomonas reinhardtii, constructed in the phage expression vector lambda gt11, was probed with antiserum directed against the nuclear-encoded gamma subunit of the chloroplast H+-transporting ATP synthase [ATP phosphohydrolase (H+-transporting) or chloroplast coupling factors 0 and 1, EC 3.6.1.34] of C. reinhardtii. A cDNA was isolated and transcribed in vitro. The transcript was translated in vitro and immunoprecipitated with anti-gamma-subunit serum to yield a product that coelectrophoresed with the immunoprecipitated product from in vitro-translated polyadenylylated RNA. These proteins were larger than the mature gamma subunit, either immunoprecipitated as chloroplast coupling factor 1 or as the individual subunit. Thus, the gamma subunit is synthesized as a precursor of greater molecular weight in C. reinhardtii. Furthermore, the precursor protein encoded by the cDNA is imported into pea chloroplasts and processed to a lower molecular weight polypeptide that coelectrophoreses with mature C. reinhardtii gamma subunit. The largest cDNA isolated is about the same length as the corresponding mRNA (approximately equal to 1900 bases long) and probably contains the entire coding region. Southern blot analyses revealed restriction fragment length polymorphisms and that the gamma subunit is probably encoded by an intron-containing single-copy gene.

Biological Transport↗

PubMed1987

Interaction of photosystem I-derived protons with the water-splitting enzyme complex. Evidence for localized domains.

The induction of millisecond delayed fluorescence mediated by PS I-dependent proton pumping has been used as an indicator of the time course with which those protons equilibrate with sites on the oxygen-evolving enzyme complex (Bowes, J. M., and Crofts, A. R. (1978). Z. Naturforsch. 33C, 271-275). We found that the induction curves were retarded by a reversible exposure of non-energized thylakoids to low concentrations of the uncoupler, desaspidin, at alkaline, but not at neutral, pH. The induction curves were not retarded by increasing the buffering capacity of the thylakoid lumen with Tricine, and were inhibited by the energy transfer inhibitors, dicyclohexylcarbodiimide (DCCD) and triphenyltin chloride (TPT). These data suggest that the catalytic site of the water-splitting complex is located in proton-sequestering membrane domains, rather than at the lumen-exposed inner membrane surface, protons released during PS I-mediated electron transport might equilibrate with protonatable sites on the oxygen-evolving complex without passing through the lumen, and those protons may travel over specific conducting pathways which can be blocked by DCCD and TPT.

Chlorophyll↗

PubMed1983

The present state of the chemiosmotic coupling theory.

Although the general principles of the chemiosmotic coupling theory have become widely accepted, the (degree of) loc(aliz)ation of electrochemical proton potential difference cannot yet be deduced from the existing experimental data. Many results are not in ready accordance with the idea that one protonic electrochemical potential difference, i.e. the one between a homogeneous inner and a homogeneous outer aqueous phase, would be the high-free-energy intermediate of membrane-linked free-energy transduction. Rather, free-energy transduction in an organelle like a mitochondrion or a chloroplast might take place in large number (about 1 per H+-ATPase) of miniature chemiosmotic systems. The energized protons produced in such a miniature system might be largely (but not totally) confined to a proton-domain belonging to it. Hence, there might be many (rather than one) different relevant proton gradients.

Chloroplasts↗