title: Substitution of murine for human CD4 residues identifies amino acids critical for HIV-gp120 binding.
abstract: Human CD4 is the receptor for the gp120 envelope glycoprotein of human immunodeficiency virus and is essential for virus entry into the host cell. Sequence analysis of CD4 has suggested an evolutionary origin from a structure with four immunoglobulin-related domains. Only the two NH2-terminal domains are required to mediate gp120 binding. The extracellular segment of murine CD4 has an overall 50% identity with its human counterpart at the amino-acid level, but fails to bind gp120. To define those residues of human CD4 critical for gp120 binding, we have taken advantage of this species difference and substituted all non-conserved murine for human CD4 residues between amino-acid positions 27-167. We used oligonucleotide-directed mutagenesis to create each of 16 individual mutant human CD4 molecules containing from 1-4 amino-acid substitutions. Introduction of as few as three amino acids into corresponding positions of human CD4 abrogates gp120 binding. Furthermore, these critical residues are located in domain I with a contribution from domain II. Modelling studies using the three-dimensional coordinates of the V kappa Bence-Jones REI homodimer localize the site in domain I to the C" beta strand within CDR2 but projecting away from the homologues of principle antigen-binding regions CDR 1 and 3.
authors.affiliation: Laboratory of Immunobiology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: L K
authors.initials: LK
authors.lastname: Clayton
authors.forename: R E
authors.initials: RE
authors.lastname: Hussey
authors.forename: R
authors.initials: R
authors.lastname: Steinbrich
authors.forename: H
authors.initials: H
authors.lastname: Ramachandran
authors.forename: Y
authors.initials: Y
authors.lastname: Husain
authors.forename: E L
authors.initials: EL
authors.lastname: Reinherz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015699:HIV Envelope Protein gp120; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D015728:Receptors, HIV; D011991:Receptors, Virus / Q000378:metabolism*; D012191:Retroviridae Proteins / Q000378:metabolism*; D014162:Transfection
chemical_list: D015699:HIV Envelope Protein gp120; D015728:Receptors, HIV; D011991:Receptors, Virus; D012191:Retroviridae Proteins