Amino acid sequence of Coprinus macrorhizus peroxidase and cDNA sequence encoding Coprinus cinereus peroxidase. A new family of fungal peroxidases

Abstract Platelet aggregation requires the binding of adhesive proteins such as fibrinogen to the heterodimer of membrane glycoproteins IIb (GPIIb) and IIIa (GPIIIa). Human erythroleukemia (HEL) cells synthesize both GPIIb and GPIIIa. Using poly(A+) RNA purified from HEL cells, we constructed a cDNA library in the lambda gt10 phage vector. This library was screened with a 38mer oligonucleotide derived from a platelet GPIIIa peptide, and three overlapping cDNAs were isolated. The three inserts encompassed 3.5 kilobases (kb), including the entire coding region of mature GPIIIa (2,286 basepairs, bp) and 1.3 kb of 3′ untranslated sequence. All 222 residues determined directly from platelet GPIIIa tryptic peptides exactly matched the HEL cell-deduced amino acid sequence. The HEL cell sequence matched a previously reported endothelial cell cDNA sequence except for eight nucleotides. Five of these nucleotide differences were silent changes consistent with genetic polymorphisms. The other three differences resulted in changes in the deduced amino acid sequence of GPIIIa; reexamination of the endothelial cell cDNA sequence in these three areas revealed that it is actually identical to the HEL cell sequence. The virtual identity of the endothelial and HEL cell cDNA sequences provides direct evidence that GPIIIa is a subunit common to cell-adhesion receptors present in more than one cell type. We localized the gene for GPIIIa to chromosome 17, the same chromosome to which we had previously mapped the gene for GPIIb.

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Amino acid sequence of flounder growth hormone deduced from a cDNA sequence

Nucleic Acids Research ◽

10.1093/nar/16.21.10362 ◽

1988 ◽

Vol 16 (21) ◽

pp. 10362-10362 ◽

Cited By ~ 14

Author(s):

Hiroshi Momota ◽

Ryo Kosugi ◽

Hideo Ohgai ◽

Akihiko Hara ◽

Hiroko Ishioka

Keyword(s):

Growth Hormone ◽

Amino Acid ◽

Amino Acid Sequence ◽

Cdna Sequence

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Peptidase E, a Peptidase Specific for N-Terminal Aspartic Dipeptides, Is a Serine Hydrolase

Journal of Bacteriology ◽

10.1128/jb.182.9.2536-2543.2000 ◽

2000 ◽

Vol 182 (9) ◽

pp. 2536-2543 ◽

Cited By ~ 16

Author(s):

Rachel A. L. Lassy ◽

Charles G. Miller

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Site Directed Mutagenesis ◽

Amino Acid Residues ◽

Serine Hydrolase ◽

Genome Sequences ◽

New Family ◽

The Family ◽

Serovar Typhimurium ◽

Structural Classes

ABSTRACT Salmonella enterica serovar Typhimurium peptidase E (PepE) is an N-terminal Asp-specific dipeptidase. PepE is not inhibited by any of the classical peptidase inhibitors, and its amino acid sequence does not place it in any of the known peptidase structural classes. A comparison of the amino acid sequence of PepE with a number of related sequences has allowed us to define the amino acid residues that are strongly conserved in this family. To ensure the validity of this comparison, we have expressed one of the most distantly related relatives (Xenopus) in Escherichia coli and have shown that it is indeed an Asp-specific dipeptidase with properties very similar to those of serovar Typhimurium PepE. The sequence comparison suggests that PepE is a serine hydrolase. We have used site-directed mutagenesis to change all of the conserved Ser, His, and Asp residues and have found that Ser120, His157, and Asp135 are all required for activity. Conversion of Ser120 to Cys leads to severely reduced (104-fold) but still detectable activity, and this activity but not that of the parent is inhibited by thiol reagents; these results confirm that this residue is likely to be the catalytic nucleophile. These results suggest that PepE is the prototype of a new family of serine peptidases. The phylogenetic distribution of the family is unusual, since representatives are found in eubacteria, an insect (Drosophila), and a vertebrate (Xenopus) but not in the Archaea or in any of the other eukaryotes for which genome sequences are available.

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Amino acid sequence of heavy chain from Xenopus laevis IgM deduced from cDNA sequence: implications for evolution of immunoglobulin domains.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.85.7.2245 ◽

1988 ◽

Vol 85 (7) ◽

pp. 2245-2249 ◽

Cited By ~ 38

Author(s):

J. Schwager ◽

C. A. Mikoryak ◽

L. A. Steiner

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Xenopus Laevis ◽

Heavy Chain ◽

Cdna Sequence

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Complete amino acid sequence of the large subunit of the low-Ca2+ -requiring form of human Ca2+ -activated neutral protease (μCANP) deduced from its cDNA sequence

FEBS Letters ◽

10.1016/0014-5793(86)80919-x ◽

1986 ◽

Vol 205 (2) ◽

pp. 313-317 ◽

Cited By ~ 108

Author(s):

Kazumasa Aoki ◽

Shinobu Imajoh ◽

Shigeo Ohno ◽

Yasufumi Emori ◽

Morio Koike ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Cdna Sequence ◽

Large Subunit ◽

Neutral Protease ◽

Complete Amino Acid Sequence

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Rhodococcus sp. Strain CR-53 LipR, the First Member of a New Bacterial Lipase Family (Family X) Displaying an Unusual Y-Type Oxyanion Hole, Similar to the Candida antarctica Lipase Clan

Applied and Environmental Microbiology ◽

10.1128/aem.06332-11 ◽

2012 ◽

Vol 78 (6) ◽

pp. 1724-1732 ◽

Cited By ~ 30

Author(s):

Arnau Bassegoda ◽

F. I. Javier Pastor ◽

Pilar Diaz

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Candida Antarctica ◽

Burkholderia Cenocepacia ◽

Sequence Motifs ◽

Oxyanion Hole ◽

Content Type ◽

New Family ◽

Consensus Amino Acid Sequence ◽

Four Blocks

ABSTRACTBacterial lipases constitute the most important group of biocatalysts for synthetic organic chemistry. Accordingly, there is substantial interest in developing new valuable lipases. Considering the lack of information concerning the lipases of the genusRhodococcusand taking into account the interest raised by the enzymes produced by actinomycetes, a search for putative lipase-encoding genes fromRhodococcussp. strain CR-53 was performed. We isolated, cloned, purified, and characterized LipR, the first lipase described from the genusRhodococcus. LipR is a mesophilic enzyme showing preference for medium-chain-length acyl groups without showing interfacial activation. It displays good long-term stability and high tolerance for the presence of ions and chemical agents in the reaction mixture. Amino acid sequence analysis of LipR revealed that it displays four unique amino acid sequence motifs that clearly separate it from any other previously described family of bacterial lipases. Using bioinformatics tools, LipR could be related only to several uncharacterized putative lipases from different bacterial origins, all of which display the four blocks of consensus amino acid sequence motifs that contribute to define a new family of bacterial lipases, namely, family X. Therefore, LipR is the first characterized member of the new bacterial lipase family X. Further confirmation of this new family of lipases was performed after cloningBurkholderia cenocepaciaputative lipase, bearing the same conserved motifs and clustering in family X. Interestingly, all lipases grouping in the new bacterial lipase family X display a Y-type oxyanion hole, a motif conserved in theCandida antarcticalipase clan but never found among bacterial lipases. This observation contributes to confirm that LipR and its homologs belong to a new family of bacterial lipases.

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Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system

Biochemical Journal ◽

10.1042/bj2300133 ◽

1985 ◽

Vol 230 (1) ◽

pp. 133-141 ◽

Cited By ~ 108

Author(s):

L P Chung ◽

D R Bentley ◽

K B Reid

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Sequence ◽

Cdna Sequence ◽

Binding Protein ◽

Regulatory Protein ◽

Protein A ◽

Classical Pathway ◽

Amino Acid Residues ◽

Cloned Cdna

By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. Immunol. 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3′ non-translated region. The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. One is at position 44, which can be glutamine or threonine, and the other is at position 309, which can be tyrosine or histidine. Northern-blot analysis indicated that the mRNA for C4b-binding protein is approx. 2.5 kilobases long. The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I.

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