Hormone-dependent processing of the avian progesterone receptor

Avian progesterone receptor exists as two forms, A and B, with molecular weights of 79,000 and 110,000 daltons, respectively. The origin and significance of these two forms is an area of active investigation and debate. Monoclonal antibodies produced against these two forms were used to examine receptor stability in cytosol and changes in the receptor forms induced by hormone binding. The lability of hormone binding at elevated temperatures is well documented. Analysis by Western blotting showed the receptor was stable in freshly prepared oviduct cytosol for 2 hr at 37°C, while hormone binding was lost within 30 min. However, loss of receptor through degradation was seen when cytosol was prepared from frozen tissue or when homogenization was excessive. Progesterone was injected into diethylstilbestrol-stimulated chicks to examine, in vivo, effects of hormone treatment on receptor forms in the cytosol and nuclear fractions. Progesterone treatment caused a time- and dose-dependent conversion of the A receptor to a form (A′) with a slower electrophoretic mobility. The cytosolic progesterone receptor was divided equally between the B and A forms, while the nuclear receptor was predominantly A′. The amount of nuclear receptor was consistently less than cytosolic receptor. Receptor phosphorylation was analyzed by incubating tissue minces with [³²P]orthophosphate with or without progesterone followed by immune isolation of receptor forms. Progesterone treatment caused a time-dependent increase in cytosol receptor phosphorylation which was evident after 5 min of treatment. This phosphorylation was observed with both the A and B receptor forms. The results indicate that receptor phosphorylation is a very early event during progesterone action.     

Author index for volume 219

Proteinase inhibitors I and II were purified to electrophoretic homogeneity from leaves of tomato plants induced by either wounding intact plants or by supplying excised plants with the proteinase inhibitor inducing factor. Affinity chromatography with chymotrypsin-Sepharose was employed as a final purification step for each inhibitor. The tomato leaf inhibitors are very similar to potato tuber inhibitors I and II in subunit molecular weight, composition, and inhibitory activities against chymotrypsin, trypsin, and subtilisin. However, unlike the potato tuber which contains multiple isoinhibitors by isoelectric focusing, the tomato leaf exhibits only two isoinhibitor forms of inhibitor I and a single form of inhibitor II. The molecular weight of native potato inhibitor I was reevaluated by rigorous ultracentrifugal analysis and compared with data from previous analyses. The data confirm that native inhibitor I has a native M_r of about 41,000 and is a pentamer. Inhibitor II has a molecular weight of near 23,000 and is a dimer.     

Use of submandibular flap to close pharyngostomes and other defects of lower anterior neck region.

We describe the use of a suprahyoid flap for reconstruction in the lower anterior neck. It has been most useful for immediately closing pharyngostomes and for resurfacing defects from the excision of squamous cell carcinoma of the larynx or other neoplasms involving the skin. This flap provides a good deal of viable tissue, it can easily be rotated without a delay procedure, and the donor area is closed by direct suture. We present our experience with 22 cases.     

Synthetic peptides including acidic clusters as substrates and inhibitors of rat liver casein kinase TS (type-2)

The hexapeptides AcSer-Glu-Glu-Glu-Val-Glu and Ser-Glu-Glu-Glu-Glu-Glu, reminiscent of the sites phosphorylated by type-2 casein kinase TS in troponin T and glycogen synthase, respectively, have been synthesized and tested as phosphorylatable substrates for casein kinase TS as well as for other protein kinases. Both peptides are readily phosphorylated by casein kinase TS but not, to any detectable extent, by either cAMP-dependent protein kinase or phosphorylase kinase. Phosphorylation by type-1 casein kinase S was almost negligible. On the other hand the hexapeptide Ser-Glu-Glu-Glu-Ala-Ala is phosphorylated much more slowly and the hexapeptide Ser-Glu-Glu-Ala-Ala-Ala is almost unaffected by casein kinase TS. While the Vmax values of casein kinase TS with the acidic hexapeptides are comparable to those obtained with the corresponding protein substrates, the apparent Km values for the peptides are about two orders of magnitude higher than those for the protein substrates. The heptapeptide Arg-Ser-Glu-Glu-Glu-Val-Glu is a very poor substrate of casein kinase TS in comparison with the corresponding hexapeptide lacking the N-terminal Arg; it is, however, a competitive inhibitor toward the protein substrates, exhibiting a Ki similar to those of Ser-Glu-Glu-Glu-Glu-Glu and (Glu)5 which, in turn, are one order of magnitude higher than that of (Glu)10. It is concluded that the minimum structural requirement of type-2 casein kinases consists of a phosphorylatable residue followed by an acidic cluster, whose length is critical for the binding to the enzyme. Additional residues on the N-terminal side are not required, but their nature can influence the transphosphorylation reaction considerably.     

Carotenoids fromCapsicum annuum fruits: Changes during ripening and storage

Contents of individual carotenoids in the fruits of Ramillete cultivar ofCapsicum annuum L. changed during ripening of recently harvested fruits or fruits stored 15 d in a dark and cool room. β-carotene and β-cryptoxanthin contents rose to a maximum and then declined, while the contents of other earotenoids rose to a rather steady level. Capsanthin was the main carotenoid which together with capsorubin were responsihle for the ripening colour.