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C T Wang

Publications and source records attributed to C T Wang.

At least 109 records · Page 6Linked to original sources
PubMed1980

Synthesis of somatostatin and [D-Trp8]-somatostatin.

Experimental details for practical syntheses of somatostatin and D-Trp8-somatostatin are described. The peptides were assembled from three fragments which permit further syntheses of analogs with modifications at positions 1, 2 or 8. N alpha-Bpoc protecting groups were used for the two major fragments and these were selectively removed in the presence of the tert.-butyl derived amino acid side chain functionalities. The two cysteine residues were protected by acetamidomethyl groups. All the peptide intermediates were fully characterized and a 10-g synthesis of the protected tetradecapeptide is reported. Major fragments were coupled by the azide method in good yield. Dihydrosomatostatin and D-Trp8-dihydrosomatostatin were isolated, purified, characterized and cyclized. Polymeric side-product was successfully recycled (by reduction with dithiothreitol and reoxidation) to give an overall yield for the oxidation of 52%. Somatostatin and D-Trp8-somatostatin were purified by gel filtration or countercurrent distribution and the final products were fully characterized and determined to be > 97% pure by reversed phase high performance liquid chromatography.

Amino Acid Sequence↗
PubMed1979

Correlation of ultrastructure of reconstituted sarcoplasmic reticulum membrane vesicles with variation in phospholipid to protein ratio.

We have previously described the reconstitution of functional membrane vesicles with lipid content similar to that of the normal sarcoplasmic reticulum membrane (approximately 1.0 mumol of phospholipid/mg of protein). The present study describes methodology to prepare reconstituted membrane vesicles with defined phospholipid to protein ratio, both lower and higher than that of the original membrane. The Ca2+ loading rate and efficiency are greatest in the membranes of highest protein content (0.38 mumol of phospholipid/mg of protein), decline slowly as the lipid content is quadrupled, and decrease markedly as the lipid content is quadrupled again. Such membranes of defined composition can be used to study lipid-protein interaction and to correlate membrane structure with composition. The number of particles observed by freeze-fracture electron microscopy can be correlated with protein content, whereas the percentage of smooth domain is proportional to the lipid content of the reconstituted membrane. Since 90% or more of the protein of the reconstituted membrane is the calcium pump protein, the number of particles observed by freeze-fracture is directly proportional to the amount of calcium pump protein in the membrane. The number of pump molecules calculated to be in the membrane is greater by a factor of two than the number of particles which we observed. This multiplicity ratio could be greater depending upon the assumptions made regarding the width of the membrane (see "Appendix"). Thus, it would appear that the particles consist of two or more molecules of pump protein. The change in protein concentration of the membrane is reflected also in thin sections and by negative staining. In thin sections, the broad inner and outer 70 A bands become discontinuous and patchy and, in the limit, approach a symmetrical 20,20,20 A trilayer as the protein content of the membrane becomes small. In an analogous fashion, the concentration of particles at the surface of the membrane, observed by negative staining, decreases with increasing lipid concentration in the membrane. Thus, the correlation of composition with structure can be observed by each of the three methods of sample preparation for electron microscopic analysis.

Adenosine Triphosphate↗
PubMed1978

Membrane asymmetry and enhanced ultrastructural detail of sarcoplasmic reticulum revealed with use of tannic acid.

Fixation of purified sarcoplasmic reticulum (SR) membrane vesicles, using glutaraldehyde supplemented with 1% tannic acid, reveals newly visualized ultrastructure in thin sections. The trilaminar appearance of the membrane is highly asymmetric; the outer electron-opaque layer is appreciably wider (70 A) than the inner layer (20 A). The asymmetry is not referable to lack of penetration of the tannic acid since: (a) SR vesicles made permeable with 1 mM EDTA, pH 8.5, show similar asymmetry; (b) treatment of SR with trypsin results in progressive loss in protein content and decrease in the thickness of the outer layer, until in the limit the trilayer has a symmetric appearance; (c) within the same muscle section, the SR membrane appears highly asymmetric whereas the sarcolemma has a more symmetric appearance; (d) reconstituted SR vesicles have a symmetric appearance with equally broad inner and outer layers (approximately 70 A); the symmetric structure is confirmed by freeze-fracture and negative staining electron microscopy. Heavy and light SR vesicles obtained by isopycnic density sedimentation of purified SR have the same asymmetric appearance of the membrane and seem to differ mainly in that the heavy vesicles contain internal contents consisting largely of Ca++-binding protein. The asymmetry of the SR membrane is referable mainly to the unidirectional alignment of the Ca++ pump protein, the major component (90% of the protein) of the membrane. The asymmetry of the SR membrane can be visualized now for the first time in situ in thin sections of muscle.

Animals↗
PubMed1977

Synthesis, biological activity, and tritiation of L-3, 4-dehydroproline-containing peptides.

A series of analogs of thyroliberin (TRH) ([L-delta3Pro3]-TRH, [D-delta3Pro3]-TRH, [L-3-MeHis2, L-delta3Pro3]-TRH) in which proline was replaced by L- or D-3, 4-dehydroproline was synthesized. The analogs exhibited approximately the same biological activity as the corresponding proline-containing peptides. These analogs and others in which 3, 4-dehydroproline is present at the NH2-terminal, COOH-terminal or internal positions in the peptide were successfully reduced with deuterium or tritium to provide the 3, 4-2H2-proline or 3,4-3H2-proline analogs, respectively, with near theoretical values of substitution. A novel procedure for the chemical resolution of DL-3, 4-dehydroproline is also described.

Animals↗
PubMed1975

Application of principles of steady-state kinetics to the in vivo estimation of acetylcholine turnover rate in mouse brain.

The rate of metabolism of acetylcholine (ACh) in mouse brain in vivo has been estimated utilizing a radio gas chromatographic approach and pulse labeling with radioactive phosphorylcholine. A kinetic model describing the change with time of the plasma choline (Ch), brain Ch and ACh specific radioactivities as interdependent variables is postulated. The experimental results are analyzed kinetically according to this model using two methods: the finite difference method described by Neff et al. (J. Pharmacol. Exp. Ther. 176: 701-710, 1971) and a simple graphic method based on the transformation of the data in terms of the decline of plasma Ch radioactivity described in this paper. The accuracy of the latter is evaluated by the classic method of the least mean squares. Calculations with the two methods yield comparable results. This method is simple and can be used to study drug effects on brain ACh turnover rate. ACh turnover rate in mouse brain has been calculated to be 0.36 mumol/g/hr with the latter method, whereas the finite difference method yields slightly lower values. Because of the compartmentalization of brain Ch and the inherent assumptions, the new method can be used only for data collected during the exponential decline of plasma radioactive choline.

Acetylcholine↗
PubMed1969

Initial ATP Formation, NADP Reduction, CO(2) Fixation, and Chloroplast Flattening Upon Illuminating Pea Leaves.

Chloroplasts in living cells of detached and sectioned leaves of Pisum sativum had a thickness of 2.68 +/- 0.04 mu in the dark as determined from photographs made using a phase contrast microscope. Upon illumination with 4000 lux for 10 min, the chloroplasts flattened to 2.15 +/- 0.04 mu. There was a short lag period of about 11 sec at 1000 lux and 2 sec at 4000 lux before appreciable light-induced flattening occurred. Both ATP and reduced nicotinamide adenine dinucleotide phosphate (NADPH) in detached pea leaves increased upon illumination and then fell during the initial 60 sec. The maximum ATP level was attained in 16 sec at 1000 lux and 10 sec at 4000 lux, while NADPH required about twice as long to reach a maximum. A sustained rate of carbon dioxide fixation occurred after a lag period coinciding in time with the drop in the NADPH level. ATP appeared to be involved not only with carbon dioxide fixation, but also with some reaction beginning sooner, perhaps the light-induced chloroplast flattening. Considering the initial photophosphorylation and the sustained CO(2) fixation rates, the ATP formation rate in vivo apparently increased after the leaves had been in the light for a few min.

Journal Article↗