Abstract
Pin1 contains an N-terminal WW domain and a C-terminal peptidyl-prolyl cis-trans isomerase (PPIase) domain connected by a flexible linker. To address the energetic and structural basis for WW domain recognition of phosphoserine (P.Ser)/phosphothreonine (P. Thr)- proline containing proteins, we report the energetic and structural analysis of a Pin1-phosphopeptide complex. The X-ray crystal structure of Pin1 bound to a doubly phosphorylated peptide (Tyr-P.Ser-Pro-Thr-P.Ser-Pro-Ser) representing a heptad repeat of the RNA polymerase II large subunit's C-terminal domain (CTD), reveals the residues involved in the recognition of a single P.Ser side chain, the rings of two prolines, and the backbone of the CTD peptide. The side chains of neighboring Arg and Ser residues along with a backbone amide contribute to recognition of P.Ser. The lack of widespread conservation of the Arg and Ser residues responsible for P.Ser recognition in the WW domain family suggests that only a subset of WW domains can bind P.Ser-Pro in a similar fashion to that of Pin1.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Binding Sites
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Crystallography, X-Ray
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Fluorescence Polarization
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Humans
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Models, Molecular
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Molecular Sequence Data
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NIMA-Interacting Peptidylprolyl Isomerase
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Peptide Fragments / chemistry
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Peptide Fragments / metabolism
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Peptidylprolyl Isomerase / chemistry*
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Peptidylprolyl Isomerase / metabolism*
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Phosphopeptides / chemistry
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Phosphopeptides / metabolism
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Phosphoserine / metabolism*
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Proline / metabolism*
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Protein Binding
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Protein Structure, Tertiary
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RNA Polymerase II / chemistry
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RNA Polymerase II / metabolism
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Sequence Alignment
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Structure-Activity Relationship
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Substrate Specificity
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Thermodynamics
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Tryptophan / metabolism*
Substances
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NIMA-Interacting Peptidylprolyl Isomerase
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Peptide Fragments
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Phosphopeptides
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Recombinant Fusion Proteins
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Phosphoserine
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Tryptophan
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Proline
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RNA Polymerase II
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PIN1 protein, human
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Peptidylprolyl Isomerase