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Biomedical subjects

A Irie

Publications and source records attributed to A Irie.

At least 91 records · Page 5Linked to original sources
PubMed1990

Glucosylceramide having a novel tri-unsaturated long-chain base from the spermatozoa of the starfish, Asterias amurensis.

Glucosylceramide (Glc beta 1-1Cer) was isolated from the spermatozoa of the starfish, Asterias amurensis. The long-chain bases of the glycolipid consisted of dihydroxy (d18:2, d18:3, d19:3, and d22:2), and trihydroxy (t22:1) types. Long-chain aldehydes derived from them were analyzed mainly by proton nuclear-magnetic resonance to determine the detailed structures. Two of the tri-unsaturated bases were identified as (4E,8E,10E)-2-amino-4,8,10-octadecatriene-1,3-di ol (d18:3) and (4E,8E,10E)-2-amino-9-methyl-4,8,10-octadecatriene+ ++-1,3-diol (d19:3), which is a novel base. Both d22:2 and t22:1 had a cis double bond at the C9 or C13 position. All fatty acids were 2-hydroxylated (C14-C25): Most of them were saturated and unbranched. About 10% was mono-unsaturated and unbranched (C22-C25), while saturated but branched (iso- and anteiso-types) C15-C18 acids were found as minor components. The main fatty acids, which summed up to more than 93% of the fatty acids in the glucosylceramide, were n-14h:0, n-15h:0, n-16h:0, n-17h:0, n-18h:0, and n-24h:1.

Aldehydes↗
PubMed1990

Gangliosides from the eggs of the sea urchin, Anthocidaris crassispina.

NeuGc alpha 2-6Glc beta 1-1Cer (M5 ganglioside) and HSO3-8NeuGc alpha 2-6Glc beta 1-1Cer (T1 ganglioside) were purified by column chromatographies with DEAE-Sephadex A-25 and silicic acid from the eggs of the sea urchin, Anthocidaris crassispina. Their chemical structures were determined by gas-liquid chromatography, methylation analysis, enzymatic hydrolysis, negative-ion fast atom bombardment mass spectrometry, and proton nuclear magnetic resonance spectroscopy. Long-chain base compositions of both gangliosides were almost identical: all the long-chain bases were phytosphingosines, and C18-phytosphingosine accounted for more than 95% of them. Fatty acid compositions were also very similar: the main fatty acids were 22:1, 23:1, 24:1, and their 2-hydroxylated forms, and the 2-hydroxy fatty acids amounted to 65.3 and 74.3% of the fatty acids in M5 and T1 gangliosides, respectively. Proton nuclear magnetic resonance spectroscopic study revealed a downfield-shifted H8 proton signal of NeuGc residue in T1 ganglioside, in agreement with the presence of sulfate ester at the C8 position.

Animals↗
PubMed1990

Ceramide dihexosides from the spermatozoa of the starfish, Asterias amurensis, consist of gentiobiosyl- cellobiosyl-, and lactosylceramide.

Ceramide dihexoside was obtained from the spermatozoa of the starfish, Asterias amurensis. Gas-liquid chromatography of the methanolysate to determine the sugar composition of the lipid demonstrated an unequal ratio of glucose and galactose, implying that two or more glycolipids are present. They were separated by thin-layer chromatography on a borate-impregnated plate into two bands. From the results of methylation analysis and chromic acid oxidation, one was determined to be lactosylceramide, while the other was suggested to be a mixture of two diglucosylceramides: gentiobiosylceramide (Glc beta 1-6Glc beta 1-1Cer) and cellobiosylceramide (Glc beta 1-4Glc beta 1-1Cer). The molar ratio of gentiobiosyl-, cellobiosyl-, and lactosylceramide was estimated to be 0.7 : 0.3 : 1.0. Ceramide dihexosides obtained from another batch of the spermatozoa, collected at the same place in a different year, consisted almost exclusively of gentiobiosylceramide as confirmed by proton-nuclear magnetic resonance spectroscopy and fast-atom bombardment mass-spectrometry. The fatty acid compositions of these glycolipids were similar and the main acids were 14h:0, 15h:0, 16h:0, 18h:0, and 24h:1 (constituting more than 80% of the total acids). The long-chain base compositions were qualitatively similar and the major constituents were commonly d18:2, d18:3, d19:3, d22:2, and t22:1. Lactosylceramide was rich in t22:1, while diglucosylceramides were rich in d22:2.

Animals↗
PubMed1989

Human urinary prokallikrein--structural analysis on activation mechanism.

The complete amino acid sequence of human urinary kallikrein has been determined. The enzyme was a single polypeptide which comprised 238 amino acid residues. In the case of prokallikrein, a propeptide which was consisted of seven amino acid residues was attached to N-terminal isoleucine of kallikrein. The sequence of Asn-X-Thr(Ser), common to glycosylation site, was identified at positions 78-80, 84-86 and 141-143. It has been shown from the sequence of kallikrein that Arg(-1)-Ile(1) and Arg(87)-Gln(88) bonds are hydrolyzed with trypsin on rapid activation of prokallikrein and the formation of disulfide-linked two chain kallikrein.

Amino Acid Sequence↗
PubMed1989

Purification and immunological properties of human urinary kallikrein and prokallikrein.

Human urinary prokallikrein and kallikrein have been purified from the same source of urine simultaneously. The anti-kallikrein and anti-prokallikrein antibodies were raised in rabbits using the purified preparations. With respect to solid phase enzyme immunoassay (EIA), immunoaffinity column chromatography, and single radial immunodiffusion, reactivity of each antibody with kallikrein was distinctly different from that with prokallikrein. Kallikrein could be determined by anti-kallikrein antibody-immobilized EIA below 20 ng per ml, whereas prokallikrein was undetectable. Prokallikrein became detectable at higher concentrations, although it was less reactive than kallikrein. The anti-prokallikrein antibody-immobilized EIA detected both kallikrein and prokallikrein with the same sensitivity. However, the binding capacity for kallikrein was about one-third less than that for prokallikrein. The results show that kallikrein in human urine may be determined directly and selectively. Similar difference in reactivity was observed with immunoaffinity column chromatography and single radial immunodiffusion. The presence of 3-4 antigenic sites per molecule was indicated by quantitative precipitin reaction, and it is suggested from analysis of amino acid sequence of kallikrein by the method of Hopp and Woods that four hydrophilic regions exist in kallikrein molecule.

Chromatography, Affinity↗
PubMed1988

Reactivities of human urinary prokallikrein and kallikrein toward rabbit antibodies with special reference to enzyme immunoassay.

Human urinary prokallikrein and kallikrein have been analyzed by means of solid phase enzyme immunoassay (EIA) using rabbit antibodies. The anti-kallikrein antibody-immobilized EIA detected both kallikrein and prokallikrein. However, only kallikrein was detectable at concentrations below 20 micrograms/1. The anti-prokallikrein antibody-immobilized EIA detected both kallikrein and prokallikrein, but the binding maximum for kallikrein was about one-third less than that for prokallikrein. Similar difference in reactivity of kallikrein and prokallikrein toward each antibody was observed with immunoaffinity column chromatography and single radial immunodiffusion. The results show that immunochemical properties of human urinary prokallikrein and kallikrein differ distinctly, and conflicting results on detectability of kallikrein and prokallikrein in human urine with EIA or radioimmunoassay using the anti-kallikrein antibody was due to the difference in reactivity of the two forms of kallikrein toward the immobilized antibody.

Animals↗
PubMed1988

Primary structure of human urinary prokallikrein.

The complete amino acid sequence of human urinary prokallikrein has been determined by amino acid analysis and sequence determination of peptide fragments obtained from chemical and enzymological cleavages of kallikrein and by comparison of the N-terminal sequence of prokallikrein with that of kallikrein, the active form. Prokallikrein was a single chain polypeptide which comprised 238 amino acid residues of kallikrein and 7 amino acid residues of the propeptide. The sequence, Asn-X-Thr(Ser), which is a common glycosylation site was found at positions 78-80, 84-86, and 141-143. Two trypsin-susceptible sites were identified. One is the Arg(-1)-Ile(1) bond and the other is the Arg (87)-Gln(88) bond. The sequence of human urinary kallikrein was identical with that of human pancreatic and kidney kallikreins (Fukushima, D. et al. (1985) Biochemistry 24, 8037-8043; Baker, A.R. & Shine, J. (1985) DNA 4, 445-459), which were predicted from the nucleotide sequences of cDNAs. The primary structure of human urinary kallikrein is homologous to those of the other animal kallikreins and kallikrein-related proteins. Key amino acid residues, His(41), Asp(96), and Ser(190), required for catalytic activity and Asp (184) required for kallikrein-type specificity are completely conserved. The results show that human urinary prokallikrein and kallikrein are of tissue type and they are excreted in urine without any modification.

Amino Acid Sequence↗
PubMed1988

Melibiosylceramide as the sole ceramide dihexoside from the eggs of the sea urchin, Anthocidaris crassispina.

Melibiosylceramide (Gal alpha 1-6Glc beta 1-1Cer) was found as the sole ceramide dihexoside from the eggs of the sea urchin, Anthocidaris crassispina. Ceramide monohexoside of the eggs consisted only of glucosylceramide (Glc beta 1-1Cer). These lipids were purified by successive column chromatographies on DEAE-Sephadex A-25, silicic acid and Florisil, and identified by gas-liquid chromatography, negative-ion fast atom bombardment mass spectrometry and proton nuclear magnetic resonance spectroscopy as well as methylation analysis. Long-chain base compositions of both lipids were almost identical and comprised n-C18-phytosphingosine and small amounts of its homologs (C17-C19). Fatty acid compositions were qualitatively very similar, but the glucosylceramide contained more 2-hydroxy fatty acid than the melibiosylceramide. Although the chain length of fatty acids was distributed over a wide range, six major fatty acids, namely 22:1, 23:1, 24:1, 22h:1, 23h:1 and 24h:1, constituted more than 92% of the fatty acid content in these lipids.

Animals↗
PubMed1987

[A clinical study of testicular tumors].

Fifty-eight of the 44,698 patients seen at our Department, between 1972 and 1984 had a testicular tumor. The incidence rate was 0.13%. The mean age of these 58 cases was 28.6 years and two peak distributions, one in the 0 to 5 year and another in 26 to 30 year age group were observed. Among them, 54 patients (93.1%) had chief complaints of painless testicular swelling at initial examination. The vast majority of them had unilateral tumors; 28 in the right and 29 in the left. Only one patient had bilateral seminomas. Histologically, 30 of them (51.7%) were seminoma, 8 were embryonal carcinoma (13.8%), 2 were teratoma (3.5%) and the remaining 18 had tumors of double or multiple histological type. Most of the seminomas were treated by a combination of high orchiectomy and radiotherapy, and chemotherapy was done mainly for non-seminomatous cases. The 5-year survival rate calculated by the Kaplan-Meier method was 100% for patients with low stage (I, II) seminomas and 62% for those with non-seminomatous tumors.

Adolescent↗
PubMed1987

Activation mechanism of human urinary prokallikrein using trypsin as a model activator.

Rapid release of a small peptide from human urinary prokallikrein by trypsin resulted in activation of the prokallikrein. The peptide was identified as the propeptide of the kallikrein from its amino acid sequence. Two large disulfide-linked peptides were also produced very slowly, which accompanied the increase in kallikrein activity. The molecular weights of the two peptides were roughly estimated to be 18,000 and 25,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). N-Terminal amino acid sequences were determined as Ile-Val-Gly-Gly-Trp-Glu-Cys-Glu-Gln-His for the Mr 18,000 peptide and Gln-Ala-Asp-Glu-Asp-Tyr-Ser-His-Asp-Leu for the Mr 25,000 peptide. The N-terminal sequence of the Mr 18,000 peptide was identical to that of the kallikrein. Both peptides contained carbohydrate side chains as judged by staining with periodic acid-Schiff's base. The results indicate strongly that trypsin hydrolyses two specific bonds of human urinary prokallikrein selectively, which are cleaved upon physiological activation to yield the two-chain kallikrein.

Enzyme Activation↗
PubMed1986

N-terminal amino acid sequence of human urinary prokallikrein.

The N-terminal amino acid sequences of human urinary prokallikrein and kallikrein have been determined. Their amino acid sequences are as follows. (Formula; see text) The results showed that prokallikrein comprises an additional seven amino acids at the amino terminus of the kallikrein, of which the sequence is (H2N)Ala-Pro-Pro-Ile-Gln-Ser-Arg(COOH). Comparison of the structure of this peptide with those of other proteins revealed extensive sequence identity with the propeptide portions of rat and mouse tissue kallikreins, that were predicted from the preproenzyme-encoded nucleotide sequences. The amino acid sequence of the peptide was also highly homologous to that of the propeptide portion of EGF-binding protein, that was predicted from the nucleotide sequence, and that of the alpha-subunit of NGF. The N-terminal amino acid sequence of kallikrein was completely identical to the reported one (Lottspeich, F., et al. (1979) Hoppe-Seyler's Z. Physiol. Chem. 360, 1947-1950) and shows considerable amino acid sequence homology with the porcine and rat pancreatic kallikreins. As far as the present results are concerned, it is strongly indicated that the inactive kallikrein in human urine is a tissue type prokallikrein which is activated on the release of the N-terminal peptide consisting of seven amino acids.

Amino Acid Sequence↗