Backbone 1H, 13C, and 15N assignments and secondary structure of bovine low molecular weight phosphotyrosyl protein phosphatase.
Phosphotyrosyl protein phosphatases play an important role in mediating cellular signal transduction; yet three-dimensional structures of this important class of proteins have not been reported. We present the sequence-specific 1H, 13C, and 15N backbone assignments for the low molecular weight bovine heart phosphotyrosyl protein phosphatase (BHPTPase) (157 residues, 17,900). The assignments were obtained from a combination of double- and triple-resonance multidimensional NMR experiments. From these assignments, the secondary structure of BHPTPase was determined from an analysis of NOE patterns, 3JHNH alpha coupling constants, 13C alpha and 13CO chemical shifts, and amide 1H exchange rates. BHPTPase was found to consist of a four-stranded parallel beta-sheet (residues K6-C12, W39-A45, Y87-M91, and K112-L116), four alpha-helices (residues I21-D32, R58-G67, S94-N104, and D135-R157), and one stretch of beta 10-helix (residues K79-F85). The secondary structure is characteristic of the beta alpha beta structural motif. The secondary structure elements identified in this study are consistent with previous chemical and mutagenesis studies of BHPTPase structure.
Zhou
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Pharmaceutical Discovery Division, Abbott Laboratories, Abbott Park, Illinois 60064.
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Thèriault
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Van Etten
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Fesik
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D000595:Amino Acid Sequence; D000818:Animals; D002247:Carbon Isotopes; D002417:Cattle; D006801:Humans; D006859:Hydrogen; D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology; D009587:Nitrogen Isotopes; D017433:Protein Structure, Secondary; D017027:Protein Tyrosine Phosphatases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D006859:Hydrogen; D017027:Protein Tyrosine Phosphatases