Triticum aestivum L. endoxylanase inhibitor (TAXI) consists of two inhibitors, TAXI I and TAXI II, with different specificities

The Triticum aestivum L. endoxylanase inhibitor (TAXI) discovered by Debyser and Delcour [(1997) Eur. Pat. filed April 1997, published as WO 98/49278] and Debyser, Derdelinckx and Delcour [(1997) J. Am. Soc. Brew. Chem. 55, 153Ő156] seems to be a mixture of two different endoxylanase inhibitors, called TAXI I and TAXI II. By using Aspergillus niger as well as Bacillus subtilis endoxylanases for assaying inhibition activity, both inhibitors could be purified to homogeneity from wheat (Triticum aestivum L., var. Soissons). TAXI I and TAXI II have similar molecular structures. They both have a molecular mass of approx. 40.0kDa, are not glycosylated and occur in two molecular forms, i.e. a non-proteolytically processed one and a proteolytically processed one. However, the pI of TAXI II (at least 9.3) is higher than that of TAXI I (8.8). TAXI I and TAXI II clearly show different inhibition activities towards different endoxylanases. The N-terminal amino acid sequences of both inhibitors show a high degree of identity, which might indicate that there is an evolutionary relationship between them.

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THE USE OF REGION SPECIFIC RADIOIMMUNOASSAYS FOR CHARACTERIZATION OF CIRCULATING CALCITONIN IN PATIENTS WITH MEDULLARY CARCINOMA OF THE THYROID GLAND

Acta Endocrinologica ◽

10.1530/acta.0.0910449 ◽

1979 ◽

Vol 91 (3) ◽

pp. 449-461 ◽

Cited By ~ 12

Author(s):

Lisbeth Myhre ◽

Kaare M. Gautvik

Keyword(s):

Amino Acid ◽

Amino Acid Sequences ◽

Molecular Forms ◽

Amino Acid Residues ◽

Terminal Amino Acid ◽

Amino Terminal ◽

Terminal Fragment ◽

Terminal Amino ◽

Carboxy Terminal ◽

Amino Terminal Fragment

ABSTRACT Two antisera with known region specificities have been used to characterize calcitonin immunoreactivity (iCT) in serum of patients with medullary thyroid carcinoma (MCT). Antiserum I which was raised against the synthetic hormone (1–32 amino acid residues), contained heterogeneous populations of immunoglobulins directed predominantly against carboxyterminal sequences of the hormone, but the antiserum reacted also with the amino-terminal fragment (1–10 amino acid residues). Antiserum II, which was raised against the carboxy-terminal hormone fragment (11–32 amino acid residues) reached equally well with the intact hormone and the C-terminal fragment, but showed negligible binding of the amino terminal fragment. Antiserum I measured therefore both amino-terminal and carboxy-terminal sequences of calcitonin while antiserum II measured only carboxy-terminal amino acid sequences. In 40 patients with MCT, antiserum I measured usually the highest concentration of serum iCT suggesting the presence of non-uniform hormone immunoreactivity. The different molecular forms of circulating iCT in 7 MCT patients were explored by using antiserum I after gel filtration on Sephadex G-100. The patients who were selected on basis of iCT measurement in serum using antiserum I and II, could be divided into 3 groups which showed characteristic iCT profiles. Group 1, in which antiserum II measured a higher concentration of serum iCT, contained predominantly (60–70 %) small fragments of calcitonin immunoreactivity. On the other hand, in the sera of group 3 in which antisera I measured an equal or the highest concentrations, the dominant form of the hormone consisted of molecular sequences equal to or larger than the intact hormone (90 %). In group 2, the two antisera measured an equal amount of serum iCT and molecular forms consisting mostly of larger hormone fragments dominated (50 %). All the patients were normocalcaemic in spite of frequently grossly elevated serum iCT, and 33 out of 36 patients had normal serum immunoreactive parathyroid hormone. In conclusion: 1. Serum iCT is heterogeneous and represents peptides of quite different molecular size with no or low biological activity. 2. Most of the serum calcitonin immunoreactivity consists of peptides with carboxy-terminal amino acid sequences. 3. Most, if not all, of the amino-terminal calcitonin immunoreactivity is due to monomeric and polymeric hormonal forms.

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Isolation of a Polygalacturonase-Inhibiting Protein (PGIP) from Wheat

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2003.16.11.955 ◽

2003 ◽

Vol 16 (11) ◽

pp. 955-961 ◽

Cited By ~ 23

Author(s):

Gabré Kemp ◽

Carl W. Bergmann ◽

Ron Clay ◽

Amie J. Van der Westhuizen ◽

Zacharias A. Pretorius

Keyword(s):

Triticum Aestivum ◽

Cell Wall ◽

Amino Acid ◽

Triticum Aestivum L ◽

Cochliobolus Sativus ◽

Terminal Amino Acid ◽

Polygalacturonase Inhibiting Protein ◽

Terminal Amino ◽

Inhibiting Protein ◽

Terminal Amino Acid Sequence

Evidence for the presence of a polygalacturonase-inhibiting protein (PGIP) from a monocotyledonous cereal is presented. A 40.3-kDa PGIP that was closely associated with the cell wall was acetone-extracted and purified from wheat (Triticum aestivum L.) leaves and stems. Wheat PGIP exhibited a highly selective inhibitory activity against endopolygalacturonase (EPG) from various fungi. Of nine EPG tested, wheat PGIP only inhibited EPG from Cochliobolus sativus, a pathogen of the tribe Poaceae. A short N-terminal amino acid sequence of wheat PGIP shows no similarity to any other characterized PGIP.

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Purification and characterization of two forms of β-d-galactosidase from rat epididymal luminal fluid: evidence for their role in the modification of sperm plasma membrane glycoprotein(s)

Biochemical Journal ◽

10.1042/bj3050041 ◽

1995 ◽

Vol 305 (1) ◽

pp. 41-50 ◽

Cited By ~ 34

Author(s):

D R P Tulsiani ◽

M D Skudlarek ◽

Y Araki ◽

M C Orgebin-Crist

Keyword(s):

Plasma Membrane ◽

Amino Acid Sequences ◽

Molecular Forms ◽

Terminal Amino Acid ◽

Sds Page ◽

Before And After ◽

Sperm Plasma Membrane ◽

Terminal Amino ◽

Ph Dependent

Previous studies from this laboratory have identified rat epididymal luminal fluid acid beta-D-galactosidase activity which also optimally hydrolyses a glycoprotein substrate at neutral pH [Skudlarek, Tulsiani and Orgebin-Crist (1992) Biochem. J. 286, 907-914]. We have now separated the luminal fluid beta-D-galactosidase into two molecular forms by ion-exchange chromatography on a column of DE-52. The separated enzyme activities were purified to an apparent homogeneity by molecular-sieve chromatography followed by affinity chromatography on a column of immobilized p-nitrophenyl beta-D-thiogalactopyranoside. The purified forms, when resolved by SDS/PAGE under reducing conditions, showed apparent molecular masses of 84 and 97 kDa. Kinetic studies, including a pH-dependent substrate preference and pH-dependent association/dissociation, disclosed no differences between these two forms. The two forms had identical N-terminal amino acid sequences. However, the 97 kDa form contained much more total carbohydrate and sialic acid than the 84 kDa form. The carbohydrate moieties in the two forms were assessed by comparing their size on SDS/PAGE before and after treatment with endo-enzymes. The removal of N-linked glycans by treatment with N-glycanase or endoglycosidase F generated de-N-glycosylated polypeptides of an apparent molecular mass of 70 kDa, and indicated that the two forms contained varying amounts of asparagine (N)-linked high mannose/hybrid-type and biantennary complex-type oligosaccharides. This result and the fact that the two molecular forms had identical N-terminal amino acid sequences indicated that the two forms probably have identical or very similar polypeptides. The potential role of the enzyme in modification of sperm plasma membrane (PM) glycoproteins was examined by resolving caput sperm PM proteins (before and after treatment in vitro of the membranes with the purified beta-D-galactosidase) on SDS/PAGE, followed by staining with peanut agglutinin (PNA), a lectin which preferentially binds to Gal beta 1,3GalNAc-linkages found in O-linked glycoproteins. The evidence presented in this report has indicated that a PNA-positive glycoprotein of an apparent molecular mass of 135-150 kDa present on the caput (but not cauda) sperm PM is degalactosylated by the digestion in vitro of the membranes with purified luminal fluid beta-D-galactosidase. This result suggests a possible role for the epididymal luminal fluid beta-D-galactosidases.

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C-Terminal amino acid sequence of chicken pepsinogen and its homology with sequences of other acid proteases

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19801144 ◽

1980 ◽

Vol 45 (4) ◽

pp. 1144-1154 ◽

Cited By ~ 3

Author(s):

Miroslav Baudyš ◽

Helena Keilová ◽

Vladimír Kostka

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

The Other ◽

Terminal Sequence ◽

Terminal Part ◽

Terminal Amino Acid ◽

Tryptic Peptides ◽

Terminal Amino ◽

High Degree ◽

Terminal Amino Acid Sequence

To determine the primary structure of the C-terminal part of the molecule of chicken pepsinogen the tryptic, chymotryptic and thermolytic digest of the protein were investigated and peptides derived from this region were sought. These peptides permitted the following 21-residue C-terminal sequence to be determined: ...Ile-Arg-Glu-Tyr-Tyr-Val-Ile-Phe-Asp-Arg-Ala-Asn-Asn-Lys-Val-Gly-Leu-Ser-Pro-Leu-Ser.COOH. A comparison of this structure with the C-terminal sequential regions of the other acid proteases shows a high degree of homology between chicken pepsinogen and these proteases (e.g., the degree of homology with respect to hog pepsinogen and calf prochymosin is about 66%). Additional tryptic peptides, derived from the N-terminal part of the zymogen molecule whose amino acid sequence has been reported before, were also obtained in this study. This sequence was extended by two residues using an overlapping peptide. An ancillary result of this study was the isolation of tryptic peptides derived from other regions of the zymogen molecule.

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Autoantibodies against triosephosphate isomerase. A possible clue to pathogenesis of hemolytic anemia in infectious mononucleosis.

Journal of Experimental Medicine ◽

10.1084/jem.171.2.565 ◽

1990 ◽

Vol 171 (2) ◽

pp. 565-570 ◽

Cited By ~ 11

Author(s):

K Ritter ◽

H Brestrich ◽

B Nellen ◽

H Kratzin ◽

H Eiffert ◽

...

Keyword(s):

Infectious Mononucleosis ◽

Triosephosphate Isomerase ◽

Clinical Symptoms ◽

Glycolytic Enzyme ◽

Amino Acid Sequences ◽

Cellular Proteins ◽

51Cr Release ◽

Terminal Amino Acid ◽

Sds Page ◽

Terminal Amino

In sera from patients with acute EBV, infection and the clinical symptoms of infectious mononucleosis antibodies of the Ig class M were found that are directed against two cellular proteins. The molecular mass of these proteins was determined to be 29 (p29) and 26 kD (p26), respectively, in SDS-PAGE. P29 was identified as part of the glycolytic enzyme triosephosphate isomerase (TPI) by comparison of the NH2-terminal amino acid sequences. A purified antibody against TPI induces a 51Cr release from human erythrocytes. Possibly, anti-TPI causes hemolysis, which is an infrequent but serious symptom of infectious mononucleosis.

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Factor D̄ of the alternative pathway of human complement. Purification, alignment and N-terminal amino acid sequences of the major cyanogen bromide fragments, and localization of the serine residue at the active site

Biochemical Journal ◽

10.1042/bj1870863 ◽

1980 ◽

Vol 187 (3) ◽

pp. 863-874 ◽

Cited By ~ 34

Author(s):

D M Johnson ◽

J Gagnon ◽

K B Reid

Keyword(s):

Amino Acid ◽

Sequence Analysis ◽

Amino Acid Sequence ◽

Alternative Pathway ◽

Amino Acid Sequences ◽

Serine Residue ◽

Amino Acid Sequence Analysis ◽

Terminal Amino Acid ◽

Terminal Amino ◽

Terminal Amino Acid Sequence

The serine esterase factor D of the complement system was purified from outdated human plasma with a yield of 20% of the initial haemolytic activity found in serum. This represented an approx. 60 000-fold purification. The final product was homogeneous as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis (with an apparent mol.wt. of 24 000), its migration as a single component in a variety of fractionation procedures based on size and charge, and its N-terminal amino-acid-sequence analysis. The N-terminal amino acid sequence of the first 36 residues of the intact molecule was found to be homologous with the N-terminal amino acid sequences of the catalytic chains of other serine esterases. Factor D showed an especially strong homology (greater than 60% identity) with rat ‘group-specific protease’ [Woodbury, Katunuma, Kobayashi, Titani, & Neurath (1978) Biochemistry 17, 811-819] over the first 16 amino acid residues. This similarity is of interest since it is considered that both enzymes may be synthesized in their active, rather than zymogen, forms. The three major CNBr fragments of factor D, which had apparent mol.wts. of 15 800, 6600 and 1700, were purified and then aligned by N-terminal amino acid sequence analysis and amino acid analysis. By using factor D labelled with di-[1,3-14C]isopropylphosphofluoridate it was shown that the CNBr fragment of apparent mol.wt. 6600, which is located in the C-terminal region of factor D, contained the active serine residue. The amino acid sequence around this residue was determined.

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Activation of bovine and chicken liver dihydrofolate reductases and its relationship to a specific cysteine residue in their NH2-terminal amino acid sequences.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)43599-5 ◽

1980 ◽

Vol 255 (14) ◽

pp. 6542-6545 ◽

Cited By ~ 3

Author(s):

B.T. Kaufman ◽

A.A. Kumar ◽

D.T. Blankenship ◽

J.H. Freisheim

Keyword(s):

Amino Acid ◽

Cysteine Residue ◽

Amino Acid Sequences ◽

Chicken Liver ◽

Terminal Amino Acid ◽

Dihydrofolate Reductases ◽

Terminal Amino

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Comparative investigations of gluten proteins from different wheat species. III. N-terminal amino acid sequences of α-gliadins potentially toxic for coeliac patients

European Food Research and Technology ◽

10.1007/s002170100365 ◽

2001 ◽

Vol 213 (3) ◽

pp. 183-186 ◽

Cited By ~ 16

Author(s):

Wieser H.

Keyword(s):

Amino Acid ◽

Amino Acid Sequences ◽

Wheat Species ◽

Terminal Amino Acid ◽

Gluten Proteins ◽

Terminal Amino

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Comparative analyses of Serratia spp. outer membrane porin proteins

Canadian Journal of Microbiology ◽

10.1139/m93-064 ◽

1993 ◽

Vol 39 (4) ◽

pp. 442-447 ◽

Cited By ~ 1

Author(s):

Joanne Hutsul ◽

Elizabeth Worobec ◽

Tom R. Parr Jr. ◽

Gerald W. Becker

Keyword(s):

Amino Acid ◽

Serratia Marcescens ◽

Single Channel ◽

Enteric Bacteria ◽

Amino Acid Sequences ◽

Chromosomal Dna ◽

Terminal Amino Acid ◽

Molecular Weight Range ◽

Terminal Amino ◽

Porin Proteins

Eight Serratia strains and several members of the Enterobacteriaceae family were used in immunoblot and Southern DNA hybridization experiments and probed with antibody and DNA probes specific for the 41-kDa Serratia marcescens porin, to determine the extent of homology between Gram-negative porins. Immunoblot analyses performed using porin-specific rabbit sera and cell envelope preparations from these strains revealed that all strains produced at least one cross-reactive protein in the 41-kDa molecular weight range. Chromosomal DNA from each of the same strains was used in Southern analyses, probed with a 20-base-length oligonucleotide probe deduced from the N-terminal amino acid sequence of the 41-kDa Serratia marcescens porin. The probe hybridized to DNA from all of the Serratia species and six of the nine other enteric bacteria. Putative porin proteins from all the Serratia species were subjected to N-terminal amino acid sequencing and porin functional analysis using the black lipid bilayer method. All amino acid sequences were identical, with one exception in which an asparagine was substituted for an aspartic acid in Serratia rubidaea. All porins had very similar porin function (single channel conductance ranging between 1.72 and 2.00 nS). The results from this study revealed that a strong conservation exists among the Serratia porins and those produced by other enteric bacteria.Key words: porins, Serratia marcescens, homology studies.

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