Sequential NMR resonance assignment and structure determination of the Kunitz-type inhibitor domain of the Alzheimer's. beta. -amyloid precursor protein

Journal Article · Tue Oct 29 00:00:00 EST 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00107a015· OSTI ID:5616215

Heald, S L; Tilton, Jr, R F ^[1]; Hammond, L J; Lee, A; Bayney, R M; Kamarck, M E; Ramabhadran, T V; Dreyer, R N; Davis, G; Unterbeck, A; Tamburini, P P ^[2]

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1. Miles, Inc., West Haven, CT (United States)
2. Molecular Therapeutics, Inc., West Haven, CT (United States)

Certain precursor proteins (APP{sub 751} and APP{sub 770}) of the amyloid {beta}-protein (AP) present in Alzheimer's disease contain a Kunitz-type serine protease inhibitor domain (APPI). In this study, the domain is obtained as a functional inhibitor through both recombinant (APPI{sub r}) and synthetic (APPI{sub s}) methodologies, and the solution structure of APPI is determined by {sup 1}H 2D NMR techniques. Complete sequence-specific resonance assignments (except for P13 and G37 NH) for both APPI{sub r} and APPI{sub s} are achieved using standard procedures. Ambiguities arising from degeneracies in the NMR resonances are resolved by varying sample conditions. Qualitative interpretation of short- and long-range NOEs reveals secondary structural features similar to those extensively documented by NMR for bovine pancreatic trypsin inhibitor (BPTI). A more rigorous interpretation of the NOESY spectra yields NOE-derived interresidue distance restraints which are used in conjunction with dynamic simulated annealing to generate a family of APPI structures. Within this family, the {beta}-sheet and helical regions are in good agreement with the crystal structure of BPTI, whereas portions of the protease-binding loops deviate from those in BPTI. These deviations are consistent with those recently described in the crystal structure of APPI.

• OSTI ID: 5616215
• Journal Information: Biochemistry; (United States), Vol. 30:43; ISSN 0006-2960
• Country of Publication: United States
• Language: English

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MOLECULAR STRUCTURE
NERVOUS SYSTEM DISEASES
OVERHAUSER EFFECT
PROTONS
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OXYGEN COMPOUNDS
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