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Analysis of the phosphorylation situation of CENP-U

ABSTRACT: Study of the reconstituted in vitro human PLK1:CENP-U complex with PBD fused N-terminally to maltose-binding-protein (MBPPBD) and residues 58-114 of CENP-U fused N-terminally to MBP (MBPCENP-U58-114). To phosphorylate T78 and T98 of MBPCENP-U58-114, we added catalytic amounts (typically 1:30 molar ratio) of active CDK1 and PLK1 kinase. Either 1) pre-phosphorylating MBPCENP-U58-114 with both kinases before adding MBPPBD for complex formation (pre-phosphorylation method); or 2) mixing MBPCENP-U58-114 and MBPPBD with both kinases for simultaneous phosphorylation and complex formation (co-mixing method) were tested.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Petra Janning

PROVIDER: MSV000101084 | MassIVE | Tue Mar 10 06:21:00 GMT 2026

SECONDARY ACCESSION(S): PXD075442

REPOSITORIES: MassIVE

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