SEIBUTSU BUTSURI KAGAKU
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
Volume 38, Issue 1
Displaying 1-8 of 8 articles from this issue
  • Yong Gang, Nobuhiko Kubo, Ikunosuke Sakurabayashi, Yoshihiko Ohtsuka, ...
    1994 Volume 38 Issue 1 Pages 1-5
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    Structural property of apolipoprotein H (β2-glycoprotein I) (apo H) was analyzed by two-dimensional electrophoresis and two-dimensional immunoblotting. Untreated purified apo H could be separated into six spots of molecular size (M=49, 000) in two-dimensional electrophoresis at pI 5-7. Spot number was decreased and size of apo H was reduced by neuraminidase treatment to three spots of molecular size 48, 000. These results suggested that structural variety of apo H was concerned partly to glycosilation of the protein. We also found that purified apo H of pooled serum from patients with hyperlipidemia has hemolytic ability to human red blood cell.
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  • Sumiko Hashimoto, Kinnya Kawano
    1994 Volume 38 Issue 1 Pages 7-12
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    Extra-band was occasionally detected in beta-fraction on cellulose acetate electrophoresis, especially when electrophoresis was done immediately after the replacement of bridging filter paper and the electrophoresis buffer. We have also noticed that the application of Toyo filter paper No. 2 as the bridging material, develops the extra-band in beta-fraction and increases the relative value of the beta-fraction, in the first run after the replacement of bridging filter paper. It has been reported that the presence of double-peak in beta-fraction on Tiselius' free electrophoresis, when electrophoresis buffer with high concentration of Ca ion was used. Therefore, we measured Ca, Mg and Fe concentrations of the eluate from two kinds of filter paper, and found relatively high concentration of Ca in the eluate from the Toyo filter paper No. 2. Immuno-fixation electrophoresis revealed that the C3 shifts from beta-fraction to slow beta-fraction and form extra-peak, when high concentration of Ca ion was exist in the electrophoresis buffer. In order to obtain more precise data on protein fractionation by cellulose acetate electrophoresis, it is essential to pay much attention on the selection of filter paper for bridging.
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  • Michinari Yokohama, Jorge Oltra C., Jorge E. Correa
    1994 Volume 38 Issue 1 Pages 13-18
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    Serum proteins of South American camelids (llama, alpaca and llamahnanaco) were identified and analyzed by microscale multisample two-dimensional (2-D) electrophoresis and immunoblotting. And then, the two-dimensional identification maps were first prepared as basic data to examine the physiological and clinico-chemical informations occurring in serum proteins. The results obtained are as follows: 1) Two-dimensional identification maps of serum proteins of the llama, alpaca and llamahnanaco were obtained in this study. 2) Serum proteins of llama, alpaca and llamahnanaco were respectively separated into 80, 83 and 78 protein spots by the 2-D electrophoresis. Serum proteins identified by immunoblotting and biochemical techniques were the following 23 components; IgM, IgG, FN, ATIII, α2M, C9, C5, C1q, C3a, C2s, BF, Cho-Es, Es, Pmg, C5, α1AT, Hp, Cp, Hpx, α1Mi, Tf, Gc and Al. 3) Unidentified three spot components ((1), (2) and (3)) detected on the separating area between Hp and Tf components are thought to be serum components to peculiar South American camelids. Functional heterogeneities caused by binding with Hb components were also observed in the Hp component. 4) Biochemical variants of Gc components were detected by the 2-D electrophoresis. The gene frequencies of A, B and C allels were 0.525, 0.050 and 0.425, respectively, in 20 South American camelids.
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  • Michinari Yokohama, Taisuke Yamazaki, Yuichi Kameyama, Yoshirou Ishiji ...
    1994 Volume 38 Issue 1 Pages 19-23
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    Physical effects of sampling conditions and preservation time on the nature of mitochondrial DNA (mtDNA) were studied by analyzing electrophoretic patterns and measuring volume and purity of mtDNA which was fractionated and extracted from organs preserved for a maximum 1.4 years in eight animal species. The analysis showed that the volume and purity of the prepared samples did not differ significantly between long-term preserved and fresh organs. Also, no physical effect of the long-term freezing preservation on the cleavage patterns was detected in the results using eight restriction endonucleases. Non-degenerate mtDNA was extracted from the livers of Japanese Sika deer which died in accidents, and the organs preserved at less than -80°C for one year. Almost no physical effect on volume or purity of the prepared mtDNA was observed. When mitochondria was fractionated from organs of animals without bleeding and with fatty livers, the non-degenerate mtDNA was obtained more by repeated washings with the homo buffer and careful cutting with scissors. From the above results, even if organs are obtained under inferior sampling conditions, as in the case of wild animals which died by traffic accidents, mtDNA obtained by fractionation can be used to study cytoplasmic inheritance. It is suggested that accident carcasses of wild animals, which are also important as gene resources can be efficiently used for academic researches.
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  • Masato Maekawa, Kayoko Sudo, Kiyotaka Fujita, Naofumi Yoshioka, Ikunos ...
    1994 Volume 38 Issue 1 Pages 25-29
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    A slow type of an electrophoretic variant of lactate dehydrogenase (LDH)-B(H) subunit was analyzed for genetic mutation. DNA analysis of the variant alleles detected a base substitution, an A to T transition at codon 320 (GAT→GTT). The mutation resulted in the replacement of an aspartic acid by valine (D 320V), and in a change of electrophoretic charge. The change may cause the net charge of the variant subunit resulting in an electrophoretic B subunit variant of the slow type. The missense mutation created a new restriction site GTNAC for endonuclease Mae III in exon 7 of the LDH-B mutant gene. Thus, we can easily detect and confirm the missense mutation.
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  • Focusing on gastrointestinal diseases
    Akira Kikuno, Masukazu Kitada, Shin-ichiro Watanabe, Tadashi Takeuchi, ...
    1994 Volume 38 Issue 1 Pages 31-36
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    Differences between alkaline phosphatase (ALP) isozyme patterns obtained by cellulose acetate membrane (CA) and polyacrylamide disc gel (PAG) electrophoreses were compared. With the PAG, high-molecular mass ALPs can be detected more frequently than those with the conventional CA. The high-molecular mass ALP migrated to the pre-liver ALP position on the CA, but was separated into the two retained ALP fractions on the PAG: one is that detected at the top position of the stacking gel (retained 2, R2); the other is that detected at a position between the stacking and the loading gels (retained 1, R1). After treatment of serum with 2% Triton X-100, the two retained ALPs could hemogeneously migrated to the position of biliary ALP. The contents of the R1 and R2 ALPs in serum were related to pathological conditions of gastrointestinal disorders. Although the R2 ALP was frequently observed in benign hepatobiliary disorders, the R1 ALP detected more in the gallbladder and pancreas carcinomas. The retained ALPs were extracted from the PAG with 2% Triton X-100 and treated with phosphatidylinositol specific phospholipase C. The resulting aliquots were further subjected to the PAG electrophoresis for analyzing the organic origin of ALP isozymes. The results showed that the R2 ALP contained the liver, bone, and intestinal ALP(s), whereas the R1 ALP had mainly liver ALP. These findings suggest that the retained ALPs serves a useful information for the diagnosis of damaged organs in clinical practice because of the organ specificity of R1 and R2 ALPs.
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  • Shoji Yamada, Mizue Inaba, Shoki Terada, Morimichi Fukuda
    1994 Volume 38 Issue 1 Pages 37-45
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
    Cobalamin-binding capacity of human seminal plasma amounted on average, to 23pmol/ml which was 20 times larger than that of serum. Most of the binding capacity was, in contrast to serum, unsaturated. Cobalamin-binders of seminal plasma separated into two distinct peaks on gel filtration; their elution positions and reactivity with antisera revealed that the largest portion of binding capacity is attributable to haptocorrin and the smaller portion to transcobalamin respectively. Total cobalamin-binding capacity and the ratio haptocorrin to transcobalamin varied significantly among samples examined. On column isoelectrofocusing, major peaks were seen at pH4.7 and 6.6; the former peak corresponding to 121 kDa reacted with an antihaptocorrin serum. One peculiar feature of the cobalamin-binder in seminal plasma is marked heterogeneity of components with pI below 4 as observed by the modified agarose-gel isoelectrofocusing technique. This suggested the presence of increased amounts of sialic acid residues on the binder molecule. The physiological role of cobalamin-binders in seminal plasma, such as the relationship to spermatogenesis, is still largely unknown. Detailed analysis of the binder may open up a new arena of cobalamin bioeffects.
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  • Toshio Okazaki, Hideji Arakawa, Matsuo Muraoka, Takashi Oda, Tatsuo Na ...
    1994 Volume 38 Issue 1 Pages 47-49
    Published: February 15, 1994
    Released on J-STAGE: March 31, 2009
    JOURNAL FREE ACCESS
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