Genetic analysis of human lymphocyte proteins by two-dimensional gel electrophoresis: 3. Frequent occurrence of genetic variants in some abundant polypeptides of PHA-stimulated peripheral blood lymphocytes

Summary The 100 or so most intensely Coomassie blue-stained polypeptides from PHA-stimulated peripheral blood lymphocytes were analyzed by two-dimensional electrophoresis in combination with family and population studies. Besides polymorphic lymphocyte cytosol 64k polypeptide reported previously, genetic variants were frequently observed in three polypeptides with molecular weights of 100,000, 49,000, and 40,000. All of them occur in the cytosol. These variant polypeptides are charge variants, because they are separated in the isoelectric focusing dimension. It is indicated by family and population studies and cell distribution analysis that the polypeptide with a molecular weight of 100,000 shows a genetic polymorphism determined by two alleles at a new autosomal locus, as described in the following paper. Family and population studies also suggest that a genetic polymorphism defined by alleles at an autosomal locus is present in each of the polypeptides with molecular weights of 49,000 and 40,000. In contrast to the previous reports of the extremely restricted genetic variability of the 100 or so most abundant fibroblast polypeptides, the present data indicate that common genetic variants are present at least in four of the 100 or so most intensely Coomassie blue-stained lymphocyte polypeptides. The result also shows that careful side-by-side comparison of two-dimensional electrophoresis patterns among both parents and their children is an effective method to detect genetic variant polypeptides.     

Epinephrine-independent production of hyperuricemia by means of hypothalamic stimulation in the conscious rat

The electrical stimulation of the ventromedial hypothalamus with 0.6 mA current produced an acute hyperuricemia associated with a rise in plasma allantoin in the conscious rat. Adrenal demedullation did not depress the hyperuricemic response. Even after the demedullation, neither propranolol nor phentolamine had any significant effects as well. These results indicate that endogeneous catecholamines are not involved in the production of the hyperuricemia; this is in sharp contrast with our previous result demonstrating the development of hyperuricemia via adrenal epinephrine by stimulating the ventromedial nucleus with 0.2 mA current. It is concluded therefore that ventromedial hypothalamic stimulation is capable of producing two different types of hyperuricemia, i.e. adrenal-epinephrine-dependent and epinephrine-independent, by varying the intensity of the stimulation current.     

Phase progress under low temperature treatment of the potassium uptake rhythm in a duckweed, Lemna gibba G3

Potassium uptake rhythm in the long-day duckweed Lemna gibbaG3 grown at 26?C disappeared at temperatures below 12?C. However,when the plants were returned to 26?C, the rhythm immediatelyrestarted from circadian time 12 with its normal wave form.Temperature steps from 20 to 30?C or from 30 to 20?C did notmodify the phase of the rhythm, although a step from 15 to 30?Cor from 30 to 15?C evoked a distortion in the wave form withoutintroducing any reproducible phase shift. Various periods of 9 or 4?C given during the subjective dayphase reduced the pace of rhythm progress by 40 or 60%, whilethose given during the subjective night phase did not. Theseresults suggest that the subjective day and night phases arethe energy charge and discharge phases for the underlying oscillator,respectively. Energy fluxes for the oscillators are brieflydiscussed. ¹Present address: National Institute for Basic Biology, Okazaki444, Japan. ²Present address: Aichi Gakuin University, Chikusa, Nagoya 464,Japan. (Received August 25, 1979; )     

Energy supply for potassium uptake rhythm in a duckweed, Lemna gibba G3

The energy relationships of the potassium uptake rhythm in aflow-medium culture of a duckweed, Lemna gibba G3, were investigated. Respiratory inhibitors or uncouplers (NaCN, 2,4-dinitrophenoland carbonyl-cyanide m-chlorophenylhydrazon) reduced the mesoror the average rate of potassium uptake, without remarkablydecreasing the amplitude of the rhythm. NaN₃, however, preferablysurpressed the amplitude at a concentration as low as 10^–6M. Both dichloro-phenyldimethylurea and the removal of CO₂ graduallydecreased the average rate of the uptake, although the rhythmicityitself was not eliminated completely in the absence of CO₂.In N₂, the average rate of uptake was reduced to zero, but rhythmwith a small amplitude survived. These observations suggested that respired photosynthates provideenergy for sustaining the average of potassium uptake and thatsome light dependent processes control the amplitude of therhythm. Exogenous sugar could make the potassium uptake rhythmappear for a relatively short time in the dark. ¹This study was carried out in the National Institute for BasicBiology and the Biological Institute, Faculty of Science, NagoyaUniversity (Received October 24, 1979; )     

The structure of pyridinoline,a collagen crosslink

Pyridinoline is an amino acid isolated from collagen and probably serves as a crosslink in collagen fiber. This compound was isolated on a large scale from bovine bone and investigated by ¹H-nmr and ¹³C-nmr spectroscopy, mass spectroscopy and chemical degradation. The structure is proposed on the basis of these data.     

In vitro construction of different oligomeric forms of lambdadv DNA and studies of their transforming activities.

Plasmid lambdadv1, which is in a dimeric form, was converted to a linear monomer duplex by the action of EcoRI restriction endonuclease that incises at a unique site in this plasmid genome. The resulting products were then joined by Escherichia coli DNA ligase to produce molecules with various oligomeric forms, and from these monomeric, dimeric, or trimeric circular molecules were purified. By transformation of cells with these DNAs, clones were obtained that carried lambdadv1 in a monomeric or dimeric form. The former type of clones have not been generated in vivo, except for one in a different host strain, and carriers of timeric or tetrameric lambdadv1's have not been obtained so far. It was observed that a considerable fraction of these oligomeric circular DNAs were converted to lower oligomers (e.g., from trimer to dimer) during transformation. The characteristics of the monomeric lambdadv1 carriers obtained were compared with those of dimeric lambdadv1 carriers. The stabilities of the plasmids of the two forms were the same. However, the monomeric plasmid carriers were less tolerant to lambdavir phage infection and perpetuated about 30% less plasmid genomes in monomer units. Furthermore, dimeric plasmid carriers appeared spontaneously and accumulated in cultures of the monomeric lambdadv1 carriers.