TTLL10 is a protein polyglycylase that can modify nucleosome assembly protein 1.
Certain proteins can undergo polyglycylation and polyglutamylation. Polyglutamylases (glutamate ligases) have recently been identified in a family of tubulin tyrosine ligase-like (TTLL) proteins. However, no polyglycylase (glycine ligase) has yet been reported. Here we identify a polyglycylase in the TTLL proteins by using an anti-poly-glycine antibody. The antibody reacted with a cytoplasmic 60-kDa protein that accumulated in elongating spermatids. Using tandem mass spectrometry of trypsinized samples, immunoprecipitated by the antibody from the TTLL10-expressing cells, we identified the 60-kDa protein as nucleosome assembly protein 1 (NAP1). Recombinant TTLL10 incorporated glycine into recombinant NAP1 in vitro. Mutational analyses indicated that Glu residues at 359 and 360 in the C-terminal part of NAP1 are putative sites for the modification. Thus, TTLL10 is a polyglycylase for NAP1.
Ikegami
Koji
K
Mitsubishi Kagaku Institute of Life Sciences (MITILS), Machida, Tokyo 194-8511, Japan.
Horigome
Daisuke
D
Mukai
Masahiro
M
Livnat
Itamar
I
MacGregor
Grant R
GR
Setou
Mitsutoshi
M
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002352:Carrier Proteins / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D018698:Glutamic Acid / Q000032:analysis; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D056490:Nucleosome Assembly Protein 1; D010453:Peptide Synthases / Q000378:metabolism*; D011099:Polyglutamic Acid / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013087:Spermatids / Q000737:chemistry
D000906:Antibodies; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C533557:NAP1L1 protein, human; C533558:Nap1l1 protein, mouse; D009687:Nuclear Proteins; D056490:Nucleosome Assembly Protein 1; C517816:TTLL5 protein, human; D011099:Polyglutamic Acid; D018698:Glutamic Acid; D010453:Peptide Synthases