Carboxypeptidase Y activity and maintenance is modulated by a large helical structure

Makino, Mai; Sahara, Takehiko; Morita, Naoki; Ueno, Hiroshi

doi:10.1002/2211-5463.12686

Cited by 3 publications

(1 citation statement)

References 30 publications

(49 reference statements)

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“…CADI method relies on the protease property of the carboxypeptidase Y (CPY), which has a very broad amino acid specificity, and is able to remove every amino acid from the carboxyl-terminus of peptides. 22 Thus, all linear peptides derived from endopeptidase (e.g., trypsin) digest are able to be broken down by CPY. In contrast, the disulfidebonded peptides can survive from the CPY treatment due to the blockage to the CPY-substrate recognition (Figure 1a).…”

Section: ■ Resultsmentioning

confidence: 99%

Carboxypeptidase Y Assisted Disulfide-Bond Identification with Linearized Database Search

Qiang

et al. 2021

Anal. Chem.

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A disulfide bond is an important protein post-translational modification and plays a key role in regulating protein oxidation status, protein structure, and stability. Analysis of a disulfide bond using mass spectrometry is challenging because there lacks an efficient method to separate the disulfide-linked peptides from a complex protein digest, and the MS data requires sophisticated interpretation. Here, we developed a novel disulfide bond identification strategy, termed as "carboxypeptidase Y assisted disulfide-bond identification (CADI)". CADI is able to significantly reduce sample complexity by depleting ∼90% of the linear peptides while keeping the disulfide-bonded peptides. Furthermore, all CADI data can be directly analyzed by widely used protein database search engines, such as Mascot and MaxQuant. Our data show that CADI is able to sensitively identify disulfide bonds in peptides and proteins. However, CADI has not yet achieved a satisfied in-depth coverage on complex mammalian cell lysates due to the limited enzymatic activity of carboxypeptidase Y and low occurrences of disulfide bonds in a proteome. Altogether, CADI is a useful method that can get disulfide-linked peptides enriched and analyzed with regular search engines. CADI holds great potentials to deepen the analysis of disulfide bond and other types of cross-linked peptides on the proteome scale.

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Section: ■ Resultsmentioning

confidence: 99%

Carboxypeptidase Y Assisted Disulfide-Bond Identification with Linearized Database Search

Qiang

et al. 2021

Anal. Chem.

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The NAC transcription factor CaNAC064 is a regulator of cold stress tolerance in peppers

et al. 2020

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Transcriptome Profiling Provides Insights Into Potential Antagonistic Mechanisms Involved in Chaetomium globosum Against Bipolaris sorokiniana

et al. 2020

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Chaetomium globosum Kunze is recognized as a potential biocontrol fungus against spot blotch of wheat caused by Bipolaris sorokiniana. Its molecular mechanism of biocontrol activity and the biosynthetic pathways involved have not been yet elucidated. Here, global transcriptome profiling of C. globosum strain Cg2 during interaction with B. sorokiniana isolate BS112 using RNA-seq was performed in order to gain insights into the potential mechanisms of antagonism. The Illumina HiSeq platform (2 × 150 bp) yielded an average of 20–22 million reads with 50–58% GC. De novo assembly generated 45,582 transcripts with 27,957 unigenes. Transcriptome analysis displayed distinct expression profiles in the interaction (Cg2–BS112), out of which 6,109 unique differentially expressed genes were present. The predominant transcripts classified as genes involved in “catalytic activity” constituted 45.06%, of which 10.02% were associated with “hydrolytic activity” (GO:0008152), and similarly, in the biological process, 29.18% of transcripts were involved in “metabolic activity” (GO:0004096 and GO:0006979). Heat map and cluster categorization suggested an increase in the expression levels of genes encoding secondary metabolites like polyketide synthase (GO:0009058), S-hydroxymethyl glutathione dehydrogenase (GO:0006069), terpene cyclase (EC 4.2.3.-), aminotran_1_2 domain-containing protein (GO:0009058), and other hydrolytic CAZYmes such as the glycosyl hydrolase (GH) family (GH 13, GH 2, GH 31, and GH 81; GO:0005975), cellulase domain-containing protein, chitinases, β-1, 3-glucanases (GO:0004565), glucan endo-1,3-beta-glucanase (GO:0052861), and proteases (GO:0004177). The obtained RNA-seq data were validated by RT-qPCR using 20 randomly chosen genes, showing consistency with the RNA-seq results. The present work is worldwide the first effort to unravel the biocontrol mechanism of C. globosum against B. sorokiniana. It generated a novel dataset for further studies and facilitated improvement of the gene annotation models in the C. globosum draft genome.

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Carboxypeptidase Y activity and maintenance is modulated by a large helical structure

Cited by 3 publications

References 30 publications

Carboxypeptidase Y Assisted Disulfide-Bond Identification with Linearized Database Search

Carboxypeptidase Y Assisted Disulfide-Bond Identification with Linearized Database Search

The NAC transcription factor CaNAC064 is a regulator of cold stress tolerance in peppers

Transcriptome Profiling Provides Insights Into Potential Antagonistic Mechanisms Involved in Chaetomium globosum Against Bipolaris sorokiniana

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