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Crystallization, sequence, and preliminary crystallographic data for an antipeptide Fab 50.1 and peptide complexes with the principal neutralizing determinant of HIV‐1 gpl20
Author(s) -
Sturam Enrico A.,
Stanfield Robyn L.,
Fieser Gail G.,
Silver Sandra,
Roguska Michael,
Hincapie L. Marina,
Simmerman Heather K. B.,
Profy Albert T.,
Wilson Ian A.
Publication year - 1992
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.340140410
Subject(s) - immunoglobulin fab fragments , peptide , orthorhombic crystal system , chemistry , crystallization , peptide sequence , monoclonal antibody , crystallography , antibody , amino acid , neutralizing antibody , stereochemistry , crystal structure , biology , biochemistry , gene , genetics , complementarity determining region , organic chemistry
X‐ray quality crystals of an Fab fragment from an antipeptide monoclonal antibody (R/V3‐50.1) that recognizes the principal neutralizing determinant (PND) of the gpl20 glycoprotein of human immunodeficiency virus type 1 (HIV‐1) (MN isolate) were grown as uncomplexed and peptide complexed forms. Crystals of the free Fab grew from high salt in orthorhombic space groups P 2 1 2 1 2 1 and I 222 and from polyethylene glycol in space groups P 1 and P 2 1 . Seeds from either the P 1 and P 2 1 native (uncomplexed) Fab crystals induced nucleation of crystals of the Fab complexed to a 16‐residue synthetic peptide corresponding to the PND when streak seeded into preequilibrated solutions of this complex. Data were collected from these complex crystals and from each of the four native Fab forms to at least 2.8Å resolution. The genes for the variable domain of the Fab were cloned and sequenced and the primary amino acid sequence was deduced from this information. Knowledge of the three‐dimensional structure of this Fab–pep‐tide complex will be important in the understanding of the PND of HIV‐1 and its recognition by neutralizing monoclonal antibodies. © 1992 Wiley‐Liss, Inc.