Mapping of fish myosin light chains by two-dimensional gel electrophoresis

1. Myosins were prepared from the ordinary muscle of 16 fish species as well as from rabbit fast muscle, and light chain subunits [alkali light chains A1, A2 and DTNB (5,5'-dithio-bis-2-nitrobenzoate) light chain] were separated on two-dimensional gel electrophoresis in combination with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2. A1 light chains showed mol. wts ranging from 21,000 to 22,900 and isoelectric points ranging from 4.51 to 4.62. DTNB light chains were spotted in a narrow area, with a mol. wt range of 16,800-17,600 and an isoelectric point range of 4.48-4.55. On the other hand, A2 light chains were most species-specific, with a mol. wt range of 14,000-19,500 and an isoelectric point range of 4.31-4.46. 3. It was suggested that the lower species-specificity in A1 as opposed to A2 is accounted for by the addition of an N-terminal peptide ("difference peptide") in the former. The properties and possible role of this peptide are discussed.     

Regulated Expression of a Gene-Fusion Product Derived from the Gene for Phytochrome I from Pisum sativum and the uidA Gene from E. coli in Transgenic Petunia hybrida

The 5'-upstream region (2.4 kb) of the gene for phytochromeI from Pisum sativum (phyl) was fused to the uidA gene fromEscherichia coli that encodes ß-glucuronidase (GUS).The resulting PHY-GUS fusion was introduced into Petunia hybridaand was used as a reporter of the expression of the phyI genewhich was recognized by GUS activity. The PHY-GUS fusion wasexpressed at a relatively high level when transgenic plantswere grown in the dark, while leaves and stems of light-grownplants showed background activity. Flowers of light-grown plantswere shown to have significant levels of GUS activity but rootsdid not have such activity. When light-grown transgenic plantswere transferred to the dark, they expressed the activity atlevels that corresponded to those of dark-grown plants. Lighttreatment prior to growth in darkness revealed red/far-red reversibilityof recovery of the activity. Thus, the 2.4-kb fragment fromthe 5' region of the phyI gene carries the information necessaryfor the light-repressible autoregulation. (Received March 30, 1991; Accepted May 20, 1991)     

Preparation and properties of the water-soluble chlorophyll-bovine serum albumin complexes

By mixing chlorophyll (Chl) a or b with a dense bovine serum albumin solution, the water-soluble Chl-bovine serum albumin complexes were prepared. These complexes, eluted near the void volume on a gel filtration, were separated well from unreacted bovine serum albumin, indicating an aggregation of such molecules in the complexes. Preparation of chlorophyllide (Chlide) a- or Chlide b-bovine serum albumin complex was unsuccessful, while the phytol-, and β-carotene-bovine serum albumin complexes could be obtained. Chls in the Chl-bovine serum albumin complexes had the following characteristics. (i) Main absorption peak of Chl a or b in the red region occurred at 675 nm or 652 nm, respectively. The Chl a-bovine serum albumin complex having absorption peak at 740 nm was also prepared. As compared with the stabilities of Chl a and b in Triton X-100. (ii) Both Chls in the bovine serum albumin-complexes were stable against oxidative stresses, such as photobleaching, Fenton reagent, peroxidase-H₂O₂ system. But (iii) they were easily hydrolyzed by chlorophyllase. These properties of Chls in the bovine serum albumin-complexes were similar to those of Chls in the isolated light-harvesting Chl a/b protein complex. A possible localization of Chls within the bovine serum albumin complexes was suggested that the porphyrin moiety of Chl was buried in bovine serum albumin; however, the hydrophilic edge of porphyrin ring, adjacent to the phytol group, occurred in the hydrophilic region of a bovine serum albumin molecule.     

Relationship between beta-lactamase activity and resistance to beta-lactam antibiotics in Mycobacterium smegmatis

Penicillin-susceptible mutants and beta-lactamase-negative mutants were isolated from Mycobacterium smegmatis after nitrosoguanidine mutagenesis. Both the mutants were found to be susceptible to low levels of penicillin and cephalosporins by twofold dilution testing. Clavulanic acid reduced the minimal inhibitory concentrations of beta-lactamase-labile beta-lactams for the penicillin-susceptible mutants and the parent strain, but had no effect on the susceptibility of the beta-lactamase-negative mutants. Comparison of the beta-lactamase activities found in these mutants and the parent strain indicated that there was a rough correlation between the beta-lactamase level in these organisms and their susceptibility to beta-lactams.     

A new species and a new subspecies of the stargazer genusGnathagnus from Northwestern Australia

Heretofore the genusGnathagnus (Uranoscopidae) has been composed of three species:G. elongatus (Temminck et Schlegel) from northern Australia to Japan,G. innotabilis (Waite) from Australasia, andG. egregius (Jordan et Thompson) from the western Atlantic Ocean. In this paper, a new species and a new subspecies from northwestern Australia are described and a key to the known species and subspecies of the genus is presented.     

Molecular Cloning and Characterization of the cDNA for Discoidin II of Dictyostelium discoideum

By using monoclonal antibodies directed against discoidin II,we have isolated cDNA clones from axenically grown Ax-2 cells.On cDNA clone (D2) condtained a 1.2-k.b insert encoding theentire discoidin II protein, which is conposed of 257 aminoacid residuces and has a calculated molecular mass of 28,574.The amino acid sequences, determined by Edman degradation ofsix tryptic peptides of discoidin II, were identical to thosededuced from the cDNA sequences. The protein bears no resemblanceto any proteins in the data banks, except that its sequenceis 49% identical with the amino acid sequence of discoidin I.Discoidin II shares with discoidin I both a carbohydratebindingsite and an Arg-Gly-Asp (RGD) sequence, which has been foundin fibronectin in mammalian cells. With the onset of aggregation(8 h of development), a 1.3-kb discoidin II mRNA begins to accumulate.A similar pattern of regulation occurs at the protein level. ¹Present address: MRC Laboratory for Molecular Cell Biology,University College London, Gower Street, London, WC1E 6BT UnitedKingdom     

Dimer Formation of a Cell-Cell Adhesion Protein, gp64 of the Cellular Slime Mold, Polysphondylium pallidum

The cellular slime mold, Polysphondylium pallidum, has two EDTA-resistanttypes of cell-cell adhesion. The major component of them hasbeen identified as a glycoprotein with a molecular mass of 64kDa on SDS-PAGE (referred to as gp64). We found that a substantialamount of the gp64 run as dimer, when gp64 was dissolved inSDS-sample buffer without 2-mercaptoethanol and then subjectedto electrophoresis. The occurrence of a homophilic dimer wasdemonstrated by analyzing the dimer-like band on a gel for itsamino acid sequence and amino acid composition. The dimer-likeband also was analyzed by three sorts of monoclonal antibodies,two of which recognize respectively a conforniational epitopeand a denatured epitope of the protein moiety of gp64. The dataindicate that the native conformation of gp64 is necessary fordimer formation. ²Present address: Institute of Immunological Science, HokkaidoUniversity, Sapporo, 060 Japan     

Food habits of sika deer on the Boso Peninsula,central japan

The rumen contents of sika deer (Cervus nippon Temminck) on the Boso Peninsula, central Japan, were analyzed to identify local, sexual and age-specific differences in food habits. Graminoids and woody plants were the primary foods throughout the year. In winter, the use of evergreen broad leaves increased. The food habits of sika deer on Boso Peninsula were intermediate between those of populations inhabiting northern and southern Japan. Acorns, mainlyLithocarpus edulis Nakai, were consumed in fall and winter with a peak in October. Since the availability of acorns is not influenced by foraging in previous years, it can be regarded as a stable food supply and hence may be important for deer on the Boso Peninsula. The local difference between the Amatsukominato (AT) area, having a large plantation ofLithocarpus producing acrons, and the Kamogawa-Katsuura (KK) area, having a small plantation ofLithocarpus, was recognized; seeds and fruit were consumed more in AT than in KK in fall and winter. Males consumed more seeds and fruit than females at both sites in fall. This can be attributed to sexual differences in nutritional requirement.