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 04:00:00 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), Journal Name: Biochemistry; (United States) Vol. 30:43; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
BARYONS
CHEMICAL SHIFT
DISEASES
ELEMENTARY PARTICLES
ENZYME INHIBITORS
FERMIONS
HADRONS
HEAVY WATER
HYDROGEN COMPOUNDS
MAGNETIC RESONANCE
MOLECULAR STRUCTURE
NERVOUS SYSTEM DISEASES
NUCLEAR MAGNETIC RESONANCE
NUCLEONS
ORGANIC COMPOUNDS
OVERHAUSER EFFECT
OXYGEN COMPOUNDS
PROTEINS
PROTONS
RESONANCE
STEREOCHEMISTRY
WATER