title: Inter-domain interactions influence the stability and catalytic activity of the bi-domain protein tyrosine phosphatase PTP99A.
abstract: The two protein tyrosine phosphatase (PTP) domains in bi-domain PTPs share high sequence and structural similarity. However, only one of the two PTP domains is catalytically active. Here we describe biochemical studies on the two tandem PTP domains of the bi-domain PTP, PTP99A. Phosphatase activity, monitored using small molecule as well as peptide substrates, revealed that the inactive (D2) domain activates the catalytic (D1) domain. Thermodynamic measurements suggest that the inactive D2 domain stabilizes the bi-domain (D1-D2) protein. The mechanism by which the D2 domain activates and stabilizes the bi-domain protein is governed by few interactions at the inter-domain interface. In particular, mutating Lys990 at the interface attenuates inter-domain communication. This residue is located at a structurally equivalent location to the so-called allosteric site of the canonical single domain PTP, PTP1B. These observations suggest functional optimization in bi-domain PTPs whereby the inactive PTP domain modulates the catalytic activity of the bi-domain enzyme.
authors.affiliation: Indian Institute of Science, Bangalore, India.
authors.forename: Lalima L
authors.initials: LL
authors.lastname: Madan
authors.forename: Kapil
authors.initials: K
authors.lastname: Goutam
authors.forename: B
authors.initials: B
authors.lastname: Gopal
mesh_terms: D000495:Allosteric Site; D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D004330:Drosophila / Q000201:enzymology; D029721:Drosophila Proteins / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D054730:Protein Interaction Domains and Motifs; D055550:Protein Stability; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry*; D054557:Receptor-Like Protein Tyrosine Phosphatases / Q000737:chemistry*
chemical_list: D029721:Drosophila Proteins; C582220:PTP99A protein, Drosophila; D017027:Protein Tyrosine Phosphatases; D054557:Receptor-Like Protein Tyrosine Phosphatases