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Detection of Nα-terminally formylated native proteins by a pan-N-formyl methionine-specific antibody.

ABSTRACT: N-formyl methionine (fMet)-containing proteins are produced in bacteria, eukaryotic organelles mitochondria and plastids, and even in cytosol. However, Nα-terminally formylated proteins have been poorly characterized because of the lack of appropriate tools to detect fMet independently of downstream proximal sequences. Using a fMet-Gly-Ser-Gly-Cys peptide as an antigen, we generated a pan-fMet-specific rabbit polyclonal antibody called anti-fMet. The raised anti-fMet recognized universally and sequence context-independently Nt-formylated proteins in bacterial, yeast, and human cells as determined by a peptide spot array, dot blotting, and immunoblotting. We anticipate that the anti-fMet antibody will be broadly used to enable an understanding of the poorly explored functions and mechanisms of Nt-formylated proteins in various organisms.

SUBMITTER: Kim D

PROVIDER: S-EPMC10164907 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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Detection of Nα-terminally formylated native proteins by a pan-N-formyl methionine-specific antibody.

Kim Dasom D Seok Ok-Hee OH Ju Shinyeong S Kim Sang-Yoon SY Kim Jeong-Mok JM Lee Cheolju C Hwang Cheol-Sang CS

The Journal of biological chemistry 20230327 5

N-formyl methionine (fMet)-containing proteins are produced in bacteria, eukaryotic organelles mitochondria and plastids, and even in cytosol. However, Nα-terminally formylated proteins have been poorly characterized because of the lack of appropriate tools to detect fMet independently of downstream proximal sequences. Using a fMet-Gly-Ser-Gly-Cys peptide as an antigen, we generated a pan-fMet-specific rabbit polyclonal antibody called anti-fMet. The raised anti-fMet recognized universally and s ...[more]

PMID: 36990220

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