Genetically encoded short peptide tag for versatile protein labeling by Sfp phosphopantetheinyl transferase.
An 11-residue peptide with the sequence DSLEFIASKLA was identified from a genomic library of Bacillus subtilis by phage display as an efficient substrate for Sfp phosphopantetheinyl transferase-catalyzed protein labeling by small molecule-CoA conjugates. We name this peptide the "ybbR tag," because part of its sequence is derived from the ybbR ORF in the B. subtilis genome. The site of Sfp-catalyzed ybbR tag labeling was mapped to the underlined Ser residue, and the ybbR tag was found to have a strong tendency for adopting an alpha-helical conformation in solution. Here we demonstrate that the ybbR tag can be fused to the N or C termini of target proteins or inserted in a flexible loop in the middle of a target protein for site-specific protein labeling by Sfp. The short size of the ybbR tag and its compatibility with various target proteins, the broad substrate specificity of Sfp for labeling the ybbR tag with small-molecule probes of diverse structures, and the high specificity and efficiency of the labeling reaction make Sfp-catalyzed ybbR tag labeling an attractive tool for expanding protein structural and functional diversities by posttranslational modification.
Yin
Jun
J
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
Straight
Paul D
PD
McLoughlin
Shaun M
SM
Zhou
Zhe
Z
Lin
Alison J
AJ
Golan
David E
DE
Kelleher
Neil L
NL
Kolter
Roberto
R
Walsh
Christopher T
CT
D000595:Amino Acid Sequence; D001412:Bacillus subtilis; D001426:Bacterial Proteins; D015335:Molecular Probes; D009842:Oligopeptides / Q000235:genetics*; D019151:Peptide Library; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism*
D001426:Bacterial Proteins; D015335:Molecular Probes; D009842:Oligopeptides; D019151:Peptide Library; D011506:Proteins; D017855:Transferases (Other Substituted Phosphate Groups); C021364:holo-(acyl-carrier-protein) synthase