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21.
Kunio Yamato I-Yih Huang Helmut Muensch Akira Yoshida Heinz-Werner Goedde Dharam P. Agarwal 《Biochemical genetics》1983,21(1-2):135-145
The usualE 1 u and atypicalE 1 a human pseudocholinesterases (acylocholine acylhydrolase, EC 3.1.1.8) were purified to homogeneity. The active-site serine residue was conjugated with diisopropyl fluorophosphate and digested with trypsin. The tryptic peptide containing the active site was isolated by gel filtration followed by two-dimensional paper chromatography and electrophoresis. The amino acid sequence of the active site peptide obtained from the usualE 1 u enzyme was found to be Gly-Glu-Ser-Ala-Gly-Ala-Ser-Ala-Val-Ser-Leu. A remarkable structural homology exists between the human and the horse enzymes in their active sites. From the difference in electrophoretic mobility of the active-site peptides obtained from the usual and atypical enzymes, the probable structure of the atypical human enzyme was deduced as Gly-His-Ser-Ala-Gly-Ala-Ser-Ala-Val-Ser-Leu. 相似文献
22.
Alkaline phosphatase from pig kidney. Microheterogeneity and the role of neuraminic acid 总被引:2,自引:0,他引:2 下载免费PDF全文
Several alkaline phosphatases (EC 3.1.3.1) could be obtained from pig kidney brush-border membrane on extraction with butan-1-ol. Three of the multiple forms were separated by DEAE-cellulose chromatography and further purified. They form a regular series with different degrees of glycosylation (mainly owing to N-acetylneuraminic acid), of charge, of molecular weight, of stability to temperature, to pH and to urea, of minimal requirement for Mg2+ and of extractability by butan-1-ol. In contrast, the detectable antigenic sites, the inhibition by amino acids and the pH-dependency of Km and Vmax. were identical for these multiple forms. On treatment with neuraminidase, the multiple forms became identical in all their properties. It was therefore concluded that the microheterogeneity of alkaline phosphatase is due to different degrees of glycosylation at polypeptide chains which appear to be otherwise identical. 相似文献
23.
Summary Using the electron microscopy and electron microscopic histochemistry the authors studied the lung alveolar epithelial cell of normal young mice.Type II cell of the alveolar epithelium has characteristically numerous osmiophilic lamellar bodies. The lamellar boies are formed in the cytoplasmic vesicle, and never originate from the mitochondrion. These bodies have abundant acid phosphatase activity in their limiting membrane therefore it is considered to be lysosomal origin, but the mitochondria have no such enzyme activity.The body which is newly formed in the cytoplasmic vesicle grows up to the large lamellar body as a result of an accumulation of the fibrous dense substance, migrates to the free margin of the type II cell of alveolar epithelium, and then is discharged into the alveolar lumen as a merocrine type secretion.Acknowledgement is given to Professor Dr. Y. Sano and Professor Dr. H. Fujita, Department of Anatomy, and Assistant Professor Dr. S. Fujita, Department of Pathology, for their kind advice and criticism. 相似文献
24.
Tadao Hashimoto Yukuo Yoshida Kunio Tagawa 《Journal of bioenergetics and biomembranes》1990,22(1):27-38
An intrinsic ATPase inhibitor inhibits the ATP-hydrolyzing activity of mitochondrial F1F0-ATPase and is released from its binding site on the enzyme upon energization of mitochondrial membranes to allow phosphorylation of ADP. The mitochondrial activity to synthesize ATP is not influenced by the absence of the inhibitor protein. The enzyme activity to hydrolyze ATP is induced by dissipation of the membrane potential in the absence of the inhibitor. Thus, the inhibitor is not responsible for oxidative phosphorylation, but acts only to inhibit ATP hydrolysis by F1F0-ATPase upon deenergization of mitochondrial membranes. The inhibitor protein forms a regulatory complex with two stabilizing factors, 9K and 15K proteins, which facilitate the binding of the inhibitor to F1F0-ATPase and stabilize the resultant inactivated enzyme. The 9K protein, having a sequence very similar to the inhibitor, binds directly to F1 in a manner similar to the inhibitor. The 15K protein binds to the F0 part and holds the inhibitor and the 9K protein on F1F0-ATPase even when one of them is detached from the F1 part. 相似文献
25.
Ping Z. Ding Kunio Kawamura James P. Ferris 《Origins of life and evolution of the biosphere》1996,26(2):151-171
The 5-phosphorimidazolide of uridine reacts on Na+-montmorillonite 22A in aqueous solution to give oligomers as long as 7 mers. The maximum chain length increases to 9 mers and the overall oligomer yield increases when 9:1 ImpU, A5 ppA mixtures react under the same conditions. The oligomer yield and maximum chain length decreases with the structure of the added pyrophosphate in the order A5 ppA>A5 ppU>U5 ppU. Structure analysis of individual oligomer fractions was performed by selective enzymatic hydrolyses followed by HPLC analysis of the products. The regioselectivity for 3,5-bond formation is 80–90% in the 9:1 ImpU, A5 ppA reaction, a percentage comparable to that observed in the 9:1 ImpA, A5 ppA reaction. Oligomerization of ImpU is inhibited by addition of dA5 ppdA, and MeppA. No oligomers containing A5 ppU were products of the 9:1 ImpU, A5 ppA reaction, a finding consistent with the simple addition of the ImpU to the A5 ppA and not the rearrangement of an ImpU-A5 ppA adduct. Concentrations of lysine or arginine which were close to that of the ImpU did not inhibit oligomer formation. Treatment of Na+-montmorillonite with 1 M arginine yielded arginine-montmorillonite, an amino acid-mineral adduct which did not catalyze ImpU oligomerization. Neither the 4–9 mers formed in the 9:1 ImpU, A5 ppA reaction nor the 4–9 mers formed by the base hydrolysis of poly(U) served as templates for the formation of oligo(A)s. 相似文献
26.
Shinichiro Nakamura Wijit Kiatipattanasakul Hiroyuki Nakayama Fumiko Ono Ippei Sakakibara Yasuhiro Yoshikawa Naoaki Goto Kunio Doi 《Journal of medical primatology》1996,25(4):294-300
Abstract: In this study, we immunohistochemically examined the several constituents of senile plaques (SPs) and cerebral amyloid angiopathy (CAA) in aged cynomolgus monkeys. Apolipoprotein E (apoE) deposited in all mature plaques and CAA, and in half of the diffuse plaques. Alpha-1-antichymotripsin (αACT) deposited in half of the mature plaques and in one third of the CAA. Amyloid precursor protein (APP), ubiquitin (Ub), and microtubule-associated protein-2 (MAP-2) accumulated in the swollen neurites of mature plaques. Glial fibrillary acidic protein (GFAP) was detected in the astrocytes and their processes surrounding the mature plaques. Tau was detected in neither the SPs nor CAA. Therefore, mature plaques involved extracellular Aβ, apoE, and αACT, and also astrocytes and swollen neurites. However, diffuse plaques involved only extracellular Aβ and apoE. Since these features, except for tau, were consistent with those in humans, this animal model will be useful for studying the pathogenesis of cerebral amyloid deposition. 相似文献
27.
Satoshi Muraki Masahiro Yamasaki Yoshito Ehara Kunio Kikuchi Kunihiro Seki 《European journal of applied physiology and occupational physiology》1996,74(5):481-483
The purpose of the present study was to examine the effect of maximal arm exercise on the skin blood circulation of the paralyzed
lower limbs in persons with spinal cord injury (PSCI). Eight male PSCI with complete lesions located between T3 and L1 performed
graded maximal arm-cranking exercise (MACE) to exhaustion. The skin blood flux at the thigh (SBFT) and that at the calf (SBFC)
were monitored using laser-Doppler flowmeter at rest and for 15 s immediately after the MACE. The subject's mean peak oxygen
uptake and peak heart rate was 1.41 ± 0.22 1·min−1 and 171.6 ± 19.2 beats·min−1, respectively. No PSCI showed any increase in either SBFT or SBFC after the MACE, when compared with the values at rest.
These results suggest that the blood circulation of the skin in the paralyzed lower limbs in PSCI is unaffected by the MACE. 相似文献
28.
Kunio Takeda Kyoko Ogawa Masami Ohara Satoshi Hamada Yoshiko Moriyama 《The protein journal》1995,14(8):679-684
Four disulfide bridges of bovineα-lactalbumin (α-lact) were selectively reduced to obtain its derivatives with three, two, and zero disulfide bridges (designated as 3SS, 2SS, and OSSα-lact, respectively). The original helicity was almost maintained in 3SSα-lact missing only the Cys6-Cysl20 bridge. Upon the reduction of both Cys28-Cys111 and Cys6-Cys120 bridges, various changes occurred in the protein. In particular, the maximum fluorescence of 1-anilinonaphthalene-8-sulfonic acid was observed in this stage. Upon the reduction of all disulfide bridges, the hydrophobic box of the protein, formed by Trp60, Ile95, Tyr103, and Trp104, was disrupted and an internal helical structure was destroyed. The conformation of each derivative was examined mainly in a solution of sodium dodecyl sulfate. In the surfactant solution, the helicity increased from 33% to 37% in 3SSα-lact, from 26% to 31% in 2SSα-lact, and from 18% to 37% in OSSα-lact, as against from 34% to 44% in intactα-lact. On the other hand, the tryptophan fluorescence of each derivative was affected in very low surfactant concentrations, suggesting that the tertiary structure considerably changed prior to the secondary structural change in the surfactant solution. 相似文献
29.
The Na,K-ATPase (sodium pump) plays a central role in the physiology of arthropod photoreceptors as it re-establishes gradients for Na+ and K+ after light stimulation. We have mapped the distribution of the Na,K-ATPase in the photoreceptors of the blowfly (Calliphora erythrocephala) by immunofluorescent and immunogold cytochemistry, and demonstrate that the distribution pattern is more complex than previously presumed. High levels of sodium pumps have been detected consistently in all photoreceptors R1-8 on the nonreceptive surface, but no sodium pumps are found on the microvillar rhabdomere. Within the nonreceptive surface of the cells R1-6, however, the sodium pumps are confined to sites juxtaposed to neighboring photoreceptor or glial cells; no sodium pumps have been detected on the parts of the nonreceptive surface exposed to the intra-ommatidial space. In R7 and R8, the sodium pumps are found over the entire nonreceptive surface. The cytoskeletal protein spectrin colocalizes with the sodium pumps suggesting that linkage of the pump molecules to the spectrin-based submembrane cytoskeleton contributes to the maintenance of the complex pattern of pump distribution. 相似文献
30.
Dopamine-Releasing Action of 6R-l-erythro-Tetrahydrobiopterin: Analysis of Its Action Site Using Sepiapterin 总被引:2,自引:1,他引:1
Abstract: Recently, we reported that 6 R - l - erythro -tetrahydrobiopterin (6 R -BH4 ), a natural cofactor for hydroxylases of tyrosine and tryptophan, has a monoamine-releasing action independent of its cofactor activity. Here we attempted to determine whether 6 R -BH4 acts inside the cell or from the outside of the cell by using brain microdialysis in the rat striatum. For this purpose, sepiapterin, an immediate precursor of 6 R -BH4 in the salvage pathway, was used to selectively increase the intracellular 6 R -BH4 levels. Dialytic perfusion of sepiapterin increased tissue levels of reduced biopterin (mainly 6 R -BH4 ) but not the extracellular levels. Administration of sepiapterin increased the extracellular levels of 3,4-dihydroxyphenylalanine (DOPA) (an index of in vivo tyrosine hydroxylase activity) and of dopamine (DA) (an index of in vivo DA release). Either of the increases was eliminated after pretreatment with a tyrosine hydroxylase inhibitor α-methyl- p -tyrosine. Administration of 6 R -BH4 increased extracellular levels of reduced biopterin, DOPA, and DA. After pretreatment with α-methyl- p -tyrosine, the increase in DOPA levels was abolished, but most of the increase in DA levels persisted. The increase in DA levels also persisted after pretreatment with nitric oxide synthase inhibitors. These data demonstrate that 6 R -BH4 stimulates DA release directly, independent of its cofactor action for tyrosine hydroxylase and nitric oxide synthase, by acting from the outside of neurons. 相似文献