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Biomedical subjects

S Odani

Publications and source records attributed to S Odani.

At least 91 records · Page 5Linked to original sources
PubMed1982

An unusual papain cleavage of a human IgG1 (lambda) myeloma protein (Mot).

An IgGl (lambda) protein which showed a unique susceptibility towards papain digestion was isolated from the serum of a patient (Mot) with multiple myeloma. The Fab fragments of this protein were degraded rapidly into smaller peptides via an Fb fragment [Gall & D'Eustachio (1972), Biochemistry 11, 4621-4628], which corresponded to the constant domains (Cl-Chl). Structural analysis of the isolated Fab fragment, which consisted of the intact L-chain, a 17,000 and a 5000 mol. wt peptide fragment, indicated that the initial cleavage site was located in the vicinity of the second hypervariable region of the Fd fragment. Examination of the partial amino acid sequences of the Mot H-chain suggested that the variable region of the H-chain may be a hitherto unknown hybrid of subgroups I and III. This particular structure seems to have made the Fab fragment highly susceptible to papain. In the course of the present study, we also found in the papain digests of several human IgG proteins an 'intermediate' 5S fragment, which had previously been reported exclusively for the papain digest of rabbit IgG.

Amino Acid Sequence↗
PubMed1982

Chicken antithrombin. Isolation, characterization, and comparison with mammalian antithrombins and chicken ovalbumin.

Chicken antithrombin was purified from fresh chicken plasma by affinity chromatography using heparin-agarose, and its amino acid and carbohydrate compositions, amino-terminal sequence, inhibition of human thrombin, and immunological properties were studied and compared with previously studied mammalian antithrombins (human, pig, rabbit, and rat), and also with chick ovalbumin. Chicken antithrombin is a single-chain glycoprotein with a total carbohydrate content of 17.5%, including 6.0% N-acetylglucosamine, 8.7% hexose, and 2.8% N-acetylneuraminic acid. The molecular weight estimated from sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis was 60,000. The amino-terminal sequence has been determined as Ala-Pro-Tyr-Ala-Val-Glu-Asp-Ile-Cys-Thr-Ala-Lys-Pro-Thr-Asp-Ile-Pro-Val-Asn, which is highly homologous to the terminal sequences of mammalian antithrombins, although the first 4 residues are quite different from those of mammalian species. Chicken antithrombin showed a stoichiometric inhibition against thrombin. The apparent dissociation constant (K1) for the complex was 6.4 X 10(-8) M. No immunological cross-reactivity was observed between chicken and mammalian antithrombins. Ovalbumin, which Hunt and Dayhoff (Biochem. Biophys. Res. Commun. 95, 864-871, 1980) proposed should be grouped in the same superfamily as antithrombin, showed neither immunological cross-reactivity with antithrombin or with its carboxymethylated derivative, nor any effect on the thrombin-antithrombin interaction. Ovalbumin showed no inhibitory effect on porcine elastase, either.

Amidohydrolases↗
PubMed1980

Importance of the carboxyl-terminal four amino acid residues in the inhibitory activity of Streptomyces subtilisin inhibitor (with a revision of its carboxyl-terminal sequence).

The carboxyl-terminal amino acid sequence of Streptomyces subtilisin inhibitor (SSI) was reinvestigated by analysis of the amino acid sequences of the thermolysin peptides from the C-terminal decapeptide, and the sequence -Val110-Ala-Phe-Phe113, which was reported in J. Biochem. 76, 1191-1209 (1974), was revised to -Val110-Phe-Ala-Phe113. Carboxypeptidase A digestion of SSI resulted in loss of the inhibitory activity in parallel with the release of the carboxyl-terminal four amino acid residues. The resulting modified inhibitor, des(Val110-Phe113)-SSI, possessed almost full inhibitory activity against subtilisin BPN' when the inhibitor was incubated with the enzyme in amounts less than one mol of enzyme per mol of the inhibitor. However, no inhibitor activity was observed when the molar ratio of the inhibitor to the enzyme was less than one. This phenomenon suggests that the carboxyl-terminal four amino acid residues might play an important role in the maintenance of the three-dimensional structure of SSI, which resists the action of the proteinase. The addition of more than 30-fold molar excess of SSI-(104-113)-decapeptide (C-terminal decapeptide of SSI) to the modified inhibitor resulted in refolding of the polypeptide chain, rendering it immune from proteolytic digestion.

Amino Acid Sequence↗
PubMed1980

The reactive site amino acid sequences of silktree (Albizzia julibrissin) seed proteinase inhibitors.

Two silktree (Albizzia julibrissin) seed proteinase inhibitors, A-II and B-II, which are homologs with soybean trypsin inhibitor (Kunitz), were cleaved with cyanogen bromide. Sequence analyses of the resulting fragments and comparison with the soybean inhibitor revealed the amino acid sequences around the reactive sites of both inhibitors. The primary reactive site of trypsin-chymotrypsin inhibitor A-II was identified as Arg66-Ile67 and that for chymotrypsin-elastase inhibitor B-II as Leu62-Met63. These results are the first reports of the reactive site sequences of homologous of soybean trypsin inhibitor (Kunitz).

Amino Acid Sequence↗
PubMed1980

Chemical substitutions of the reactive site leucine residue in soybean Bowman-Birk proteinase inhibitor with other amino acids.

A method of exchanging amino acid residue (P1 residue) which determines primarily the inhibitory specificity of a proteinase inhibitor is described. Leu43 (P1 residue) at the chymotrypsin reactive site of Bowman-Birk soybean inhibitor was removed by limited proteolysis of Leu43-Ser44 bond followed by carboxypeptidase digestion. An appropriate amino acid methyl ester was introduced into this position by water-soluble carbodiimide. Derivatives having methyl esters of Gly, Ala, Val, Met, Leu, Phe, Trp, or D-Trp at position 43 were prepared and were shown to restore the activity to various extents. This method provides new approach to study structure-function relationship of proteinase inhibitors and to prepare novel inhibitors.

Amino Acids↗
PubMed1979

Characterization of a peptide released during the reaction of human alpha 1-antitrypsin and bovine alpha-chymotrypsin.

The release of a peptide (molecular weight: about 3,600) was observed during complex formation between human alpha 1-antitrypsin (alpha 1-AT) and bovine alpha-chymotrypsin, when monitored by gel-electrophoresis in the presence of sodium lauryl sulfate. Release of the peptide was proportional to the extent of complex formation. Peptides of the same molecular weight were also released during the complex formation of alpha 1-AT with bovine trypsin or porcine elastase. The peptide released from the complex with bovine alpha-chymotrypsin was composed of 32 amino acid residues, which did not correspond to the composition of any 32 amino acid segment in the bovine alpha-chymotrypsin sequence. The N- and C-terminal sequences of the peptide were determined to be H-(Ser)-Ile-Pro-Pro-Glu- and -Gln-Lys-OH, respectively. Though there was some uncertainty as to the N-terminal sequence, it is quite different from that of the original alpha-AT molecule, and showed a similarity to the sequences of the leaving group sides of the reactive sites in some legume proteinase inhibitors. The C-terminal 2 residues were identical with those of native alpha 1-AT. These results suggest that the peptide was released from the C-terminal region of alpha 1-AT uon interaction with alpha-chymotrypsin. It is tempting to suggest that alpha 1-AT inhibits a serine proteinase by the acyl enzyme mechanism at a residue adjacent to the amino group of the N-terminus of this peptide and that this peptide is liberated as a leaving group in the enzymic process.

Amino Acid Sequence↗
PubMed1979

Proteinase inhibitors from a mimosoideae legume, Albizzia julibrissin. Homologues of soybean trypsin inhibitor (Kunitz).

Four proteinase inhibitors, A-II, A-III, B-I, and B-II, were isolated from seeds of Albizzia julibrissin (silk tree) of the subfamily Mimosoideae, which is often regarded as the most primitive group of the Leguminosae plants. They were all of the high-molecular weight type (21,600 for A-II and A-III, and 19,000 for B-I and B-II), and composed of two polypeptide chains, linked together by a disulfide bond. A-II (A-III) inhibited bovine trypsin and alpha-chymotrypsin probably at an identical site. B-I (BII) inactivated bovine alpha-chymotrypsin and porcine elastase. Sequence analyses of A-II and B-II revealed a considerable homology with soybean trypsin inhibitor (Kunitz) but suggested the presence of an about 20-amino acid insertion in the A-II molecule.

Amino Acid Sequence↗
PubMed1978

Studies on soybean trypsin inhibitors, XII. Linear sequences of two soybean double-headed trypsin inhibitors, D-II and E-I.

Soybean inhibitor D-II is an inhibitor of bovine trypsin. Sequence analysis was carried out on the reduced and S-carboxymethylated protein by conventional methods to establish the complete amino acid sequence. The sequence of D-II indicated high homology with other legume inhibitors, but it was unique because of the occurrence of identical residues (arginine) at both of the reactive sites. This structure is thought to reflect that of a prototype double-headed inhibitor. The possible evolutionary process of the legume double-headed inhibitors is discussed on this basis. Comparison with another soybean inhibitor C-II suggested that a single methionine (C-II)-glutamine (D-II) replacement at the P2'position resulted in the loss of alpha-chymotrypsin inhibitory activity of D-II. The results of a hydrogen peroxide oxidation experiment on C-II supported this suggestion. The sequence of the amino-terminal 21 residues of inhibitor E-I was determined using a sequentor. It was shown that this inhibitor lacks the amino-terminal nine residues of D-II.

Amino Acid Sequence↗
PubMed1978

Studies on soybean trypsin inhibitors. XIII. Preparation and characterization of active fragments from Bowman-Birk proteinase inhibitor.

Soybean Bowman-Birk inhibitor, a double-headed inhibitor of trypsin and alpha-chymotrypsin, was treated with cyanogen bromide and then pepsin to yield two inhibitory active fragments. Structural investigation showed that one of the fragments was derived from the trypsin inhibitory domain and the other from the chymotrypsin inhibitory domain of the inhibitor. In contrast to the unusual stability of the native inhibitor, the separated domains were less stable and could be inactivated with excess proteinases. These results suggest that the legume double-headed inhibitors acquired their unusual stability by duplicating an ancestral single-headed structure.

Amino Acid Sequence↗
PubMed1977

Studies on soybean trypsin inhibitors. X. Isolation and partial characterization of four soybean double-headed proteinase inhibitors.

Four Bowman-Birk type double-headed inhibitors (B, C-II, D-II, and E-I) were isolated from soybeans. Inhibitor B was different from Bowman-Birk inhibitor only in chromatographic behavior. One mole of C-II inhibited one mole each of bovine trypsin and bovine alpha-chymotrypsin, probably at the same site, and porcine elastase at another reactive site. In the ordinary assay system D-II and E-I inhibited only trypsin activity at a non-stoichiometric inhibitor-enzyme ratio of 1:1.4, and the complexes had rather high dissociation constants. These inhibitors were all inactive toward subtilisin BPN'.

Amino Acids↗