Characterization of the sulfated glycopeptide of chicken pepsinogen

S-sulfonated chicken pepsinogen was digested with TPCK-trypsin; large tryptic peptides, separated on Sephadex G-25 fine, were subjected to additional cleavage with α-chymotrypsin. The hold-up fraction of the chymotryptic digest from the Sephadex G-25 column, was resolved by high voltage electrophoresis. The three most acidic zones contained glycopeptides of identical amino acid sequence Val-Ser-Thr-Asn-Glu-Thr-Val-Tyr, yet differed in the composition of the sugar moiety. These glycopeptides, moreover, bear different numbers of sulfate groups which enabled the resolution of the peptides. The most acidic glycopeptide contains 7 glucosamine residues, 3 mannose residues and 5 sulfate groups, the second one 6 glucosamine residues, 3 mannose residues and 4 sulfate groups and the slowest, minority glycopeptide, 5 glucosamine residues, 2 mannose residues and 2 sulfate groups. The entire sugar moiety is attached to one of the chain viaasparagine. In other experiments the glycopeptides were also isolated from the thermolytic digest of chicken pepsin; their C-terminal sequence was shorter by two amino acid residues. The tentative assignment of the glycopeptides to the amino acid sequence of pepsinogen resulted from the analysis of the limited tryptic digest of the whole protein molecule. Chicken pepsinogen is glycosylated at the site of the chain occupied by a phosphoserine residue in hog pepsinogen A.

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Hyaluronan-binding region of aggrecan from pig laryngeal cartilage. Amino acid sequence, analysis of N-linked oligosaccharides and location of the keratan sulphate

Biochemical Journal ◽

10.1042/bj2860761 ◽

1992 ◽

Vol 286 (3) ◽

pp. 761-769 ◽

Cited By ~ 31

Author(s):

F P Barry ◽

J U Gaw ◽

C N Young ◽

P J Neame

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Sequence ◽

Signal Peptidase ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Binding Region ◽

Keratan Sulphate ◽

Laryngeal Cartilage ◽

Hyaluronan Binding

The hyaluronan-binding region (HABR) was prepared from pig laryngeal cartilage aggrecan and the amino acid sequence was determined. The HABR had two N-termini: one N-terminal sequence was Val-Glu-Val-Ser-Glu-Pro (367 amino acids in total), and a second N-terminal sequence (Ala-Ile-Ser-Val-Glu-Val; 370 amino acids in total) was found to arise due to alternate cleavage by the signal peptidase. The N-linked oligosaccharides were analysed by examining their reactivity with a series of lectins. It was found that the N-linked oligosaccharide on loop A was of the mannose type, while that on loop B was of the complex type. No reactivity was detected between the N-linked oligosaccharide on loop B' and any of the lectins. The location of keratan sulphate (KS) in the HABR was determined by Edman degradation of the immobilized KS-containing peptide. The released amino acid derivatives were collected and tested for the presence of epitope to antibody 5-D-4. On the basis of 5-D-4 reactivity and sequencing yields, the KS chains are attached to threonine residues 352 and 357. There is no KS at threonine-355. This site is not in fact in G1, but about 16 amino acid residues into the interglobular domain. Comparison of the structure of the KS chain from the HABR and from the KS domain of pig laryngeal cartilage aggrecan was made by separation on polyacrylamide gels of the oligosaccharides arising from digestion with keratanase. Comparison of the oligosaccharide maps suggests that the KS chains from both parts of the aggrecan molecule have the same structure.

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Isolation and characterization of a cDNA clone coding for the calcium-binding subunit of calcineurin from bovine brain: An identical amino acid sequence to the human protein

DNA Sequence ◽

10.3109/10425179409020857 ◽

1994 ◽

Vol 4 (5) ◽

pp. 313-318 ◽

Cited By ~ 3

Author(s):

Cheryl E. Nargang ◽

Drell A. Bottorff ◽

Kazuo Adachi

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Cdna Clone ◽

Calcium Binding ◽

Bovine Brain ◽

Human Protein ◽

Isolation And Characterization ◽

Identical Amino Acid Sequence ◽

Binding Subunit

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Amino acid sequence of penicillopepsin. IV. Myxobacter AL-1 protease II and Staphylococcus aureus protease fragments and homology with pig pepsin and chymosin

Canadian Journal of Biochemistry ◽

10.1139/o76-128 ◽

1976 ◽

Vol 54 (10) ◽

pp. 902-914 ◽

Cited By ~ 19

Author(s):

Anne Cunningham ◽

Hsin-Min Wang ◽

Stephen R. Jones ◽

Alexander Kurosky ◽

Leticia Rao ◽

...

Keyword(s):

Amino Acids ◽

Staphylococcus Aureus ◽

Amino Acid ◽

Amino Acid Sequence ◽

Sequence Data ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Fungal Enzyme ◽

Fungal Protease ◽

And Staphylococcus Aureus

The digest of penicillopepsin (EC 3.4.23.7) with protease II from Myxobacter AL-1 gave five fragments which were separated on a Biogel P-100 column in 70% formic acid. The fragments were from 16 to 125 amino acids long. Two fragments were also isolated from a digest with a protease from Staphylococcus aureus. The analysis of these fragments by automatic sequencer gave a number of overlaps of the chymotryptic and thermolytic peptides. The available amino acid sequence data for penicillopepsin described in this paper and the accompanying papers (Kurosky, A. &Hofmann, T.: Can. J. Biochem. 54, 872 (1976); Rao, L. &Hofmann, T.: Can. J. Biochem. 54, 885 (1976); Harris, C. I., Rao, L., Shutsa, P., Kurosky, A. &Hofmann, T.: Can. J. Biochem. 54, 895 (1976)) have been combined and yield 15 fragments which range in lengths from 3 to 112 amino acid residues. These unique fragments account for virtually all the amino acids of the fungal protease. Four of the fragments with a total of 194 residues (about 60% of the molecule) have been aligned with corresponding sections of pig pepsin (EC 3.4.23.1) and with part of the N-terminal sequence available for calf chymosin (EC 3.4.23.4). In the alignments about 37% of the residues in the fungal enzyme are identical with at least one of the mammalian enzymes. An additional 20% are chemically similar. These results, together with previously reported active-site directed modifications, show conclusively that penicillopepsin is an evolutionary homologue of the mammalian acid proteases.

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The amino acid sequence of ferredoxin from Brassica napus (rape)

Biochemical Journal ◽

10.1042/bj1850239 ◽

1980 ◽

Vol 185 (1) ◽

pp. 239-243 ◽

Cited By ~ 6

Author(s):

I Takruri ◽

D Boulter

Keyword(s):

Amino Acid ◽

Brassica Napus ◽

Amino Acid Sequence ◽

Polypeptide Chain ◽

Amino Acid Residues ◽

Plant Type ◽

Single Polypeptide Chain ◽

N Terminus ◽

Single Polypeptide

The amino acid sequence of the ferredoxin of Brassica napus was determined by using a Beckman 890C sequencer in combination with the characterization of peptides obtained by tryptic and chymotryptic digestion of the protein; some peptides were subdigested with thermolysin. The molecule consists of a single polypeptide chain of 96 amino acid residues and has an unblocked N-terminus. The primary structure shows considerable similarity with other plant-type ferredoxins.

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Cloning and expression of a novel Mu class murine glutathione transferase isoenzyme

Biochemical Journal ◽

10.1042/bj20020041 ◽

2002 ◽

Vol 366 (3) ◽

pp. 817-824 ◽

Cited By ~ 17

Author(s):

Jianxia GUO ◽

Ludwika ZIMNIAK ◽

Piotr ZIMNIAK ◽

John L. ORCHARD ◽

Shivendra V. SINGH

Keyword(s):

Small Intestine ◽

Amino Acid ◽

Amino Acid Sequence ◽

Glutathione Transferase ◽

Cloning And Expression ◽

Pcr Analysis ◽

Amino Acid Residues ◽

Reverse Transcription Pcr ◽

Mouse Small Intestine

The present study describes the cDNA cloning, expression and characterization of a novel Mu class murine glutathione transferase (GST) isoenzyme. Screening of a cDNA library from the small intestine of a female A/J mouse using consensus probes derived from Mu class murine GST genes (mGSTM1—mGSTM5) resulted in the isolation of a full-length cDNA clone of a previously unknown Mu class GST gene (designated as mGSTM7). The choice of tissue was based on our previous identification in female A/J mouse small intestine of a potentially novel Mu class GST isoenzyme. The deduced amino acid sequence of mGSTM7, which comprises of 218 amino acid residues, exhibited about 67—78% identity with other Mu class murine GSTs. Recombinant mGSTM7-7 cross-reacted with anti-(GST Mu) antibodies, but not with anti-(GST Alpha) or anti-(GST Pi) antibodies. The pI and the reverse-phase-HPLC elution profile of recombinant mGSTM7-7 were different from those of other Mu class murine GSTs. The substrate specificity of mGSTM7-7 was also different compared with other Mu class murine GSTs. Interestingly, mGSTM7 had a higher identity with the human Mu class isoenzyme hGSTM4 (87% identity and 94% similarity in the amino acid sequence) than with any of the known mouse Mu class GSTs. Specific activities of recombinant mGSTM7-7 and human GSTM4-4 were comparable towards several substrates. For example, similar to hGSTM4-4, recombinant mGSTM7-7 was poorly active in catalysing the GSH conjugation of 1-chloro-2,4-dinitrobenzene and ethacrynic acid, and lacked activity towards 1,2-dichloro-4-nitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane. These results suggested that hGSTM4-4 might be the human counterpart of mouse GSTM7-7. Reverse transcription-PCR analysis using mGSTM7-specific primers revealed that mGSTM7 is widely expressed in tissues of female A/J mice, including liver, forestomach, lung, kidney, colon and spleen.

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Purification and characterization of three distinct types of phospholipase A2 inhibitors from the blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus

Biochemical Journal ◽

10.1042/bj3250527 ◽

1997 ◽

Vol 325 (2) ◽

pp. 527-531 ◽

Cited By ~ 55

Author(s):

Naoki OHKURA ◽

Hiroaki OKUHARA ◽

Seiji INOUE ◽

Kiyoshi IKEDA ◽

Kyozo HAYASHI

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Phospholipase A2 ◽

Blood Plasma ◽

Amino Acid Residues ◽

Group Iii ◽

Group I ◽

Binding Surface ◽

Group Ii

Three distinct types of phospholipase A2 (PLA2) inhibitory proteins (PLIα, PLIβ, and PLIγ) were isolated from the blood plasma of the Chinese mamushi, Agkistrodonblomhoffiisiniticus. PLIα is an inhibitor that we have already purified and whose amino acid sequence we have already determined [Ohkura, Inoue, Ikeda and Hayashi (1993) J. Biochem. (Tokyo) 113, 413–419]. It inhibited selectively the group-II acidic PLA2s from Crotalidae venom. PLIβ was a 160-kDa glycoprotein having a trimeric structure composed of 50-kDa subunits. The amino acid sequence of the first 30 amino acids of the N-terminal part of the 50-kDa subunit was determined and found to have no significant homology to that of known proteins. PLIβ was a selective inhibitor against the group-II basic PLA2s from Crotalidae venom. Some amino acid residues located in or close to the interfacial binding surface of the group-II basic PLA2s were suggested to be involved in selective binding to PLIβ. PLIγ was a 100-kDa glycoprotein containing 25-kDa and 20-kDa subunits and inhibited all of the PLA2s investigated equally, including Elapidae venom PLA2s (group I), Crotalidae and Viperidae venom PLA2s (group II) and honey-bee PLA2 (group III). From the N-terminal sequences of the two subunits, PLIγ was found to be the same type of PLI that had been purified from Thailand cobra plasma.

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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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Amino acid sequence of cyanogen bromide fragment CB5 of human hemopexin

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19890803 ◽

1989 ◽

Vol 54 (3) ◽

pp. 803-810 ◽

Cited By ~ 1

Author(s):

Ivan Kluh ◽

Ladislav Morávek ◽

Manfred Pavlík

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Acid Hydrolysis ◽

Cyanogen Bromide ◽

Polypeptide Chain ◽

Amino Acid Residues ◽

Mild Acid Hydrolysis ◽

Methionine Residue ◽

Cyanogen Bromide Fragment ◽

Mild Acid

Cyanogen bromide fragment CB5 represents the region of the polypeptide chain of hemopexin between the fourth and fifth methionine residue (residues 232-352). It contains 120 amino acid residues in the following sequence: Arg-Cys-Ser-Pro-His-Leu-Val-Leu-Ser-Ala-Leu-Thr-Ser-Asp-Asn-His-Gly-Ala-Thr-Tyr-Ala-Phe-Ser-Gly-Thr-His-Tyr-Trp-Arg-Leu-Asp-Thr-Ser-Arg-Asp-Gly-Trp-His-Ser-Trp-Pro-Ile-Ala-His-Gln-Trp-Pro-Gln-Gly-Pro-Ser-Ala-Val-Asp-Ala-Ala-Phe-Ser-Trp-Glu-Glu-Lys-Leu-Tyr-Leu-Val-Gln-Gly-Thr-Gln-Val-Tyr-Val-Phe-Leu-Thr-Lys-Gly-Gly-Tyr-Thr-Leu-Val-Ser-Gly-Tyr-Pro-Lys-Arg-Leu-Glu-Lys-Glu-Val-Gly-Thr-Pro-His-Gly-Ile-Ile-Leu-Asp-Ser-Val-Asp-Ala-Ala-Phe-Ile-Cys-Pro-Gly-Ser-Ser-Arg-Leu-His-Ile-Met. The sequence was derived from the data on peptides prepared by cleavage of fragment CB5 by mild acid hydrolysis, by trypsin and chymotrypsin.

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