Preparation and crystallization of a human immunodeficiency virus p24-Fab complex

Andrew J. Prongay, Thomas Smith, Michael G. Rossmann, Lorna S. Ehrlich, Carol A. Carter, Jan McClure

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

A recombinant form of human immunodeficiency virus capsid protein, p24, expressed in Escherichia coli has been purified to homogeneity and separated into distinct isoelectric forms. A monoclonal antibody, mAb25.4, which recognizes an epitope in the amino-terminal region of p24, has been purified to homogeneity from ascites fluid and digested with papain to produce the respective antigen-binding fragment (Fab). The Fab25.4 was purified from the digestion mixture and separated into two distinct isoelectric forms. The two Fab species were each complexed with one isoelectric form of the recombinant p24 by incubating equimolar quantities of the two proteins. Two different crystal morphologies of the p24-Fab25.4 complex were obtained by the vapor-diffusion method with 12-24% PEG 3350 as the precipitant. One of these crystal forms has unit-cell parameters of a = 92.1 Å, b = 85.4 Å, c = 54.0 Å, a = γ = 90.0° and β = 90.4° and belongs to the monoclinic space group P21, with one molecule of the complex per asymmetric unit. These crystals strongly diffracted x-rays to at least 2.7-Å resolution.

Original languageEnglish (US)
Pages (from-to)9980-9984
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number24
StatePublished - 1990
Externally publishedYes

Fingerprint

Human Immunodeficiency Virus Proteins
Immunoglobulin Fab Fragments
Papain
Capsid Proteins
Crystallization
Ascites
Epitopes
Digestion
Monoclonal Antibodies
X-Rays
HIV
Escherichia coli
Antigens
Proteins
polyethylene glycol 3350

Keywords

  • AIDS
  • Capsid protein
  • Crystallography
  • Monoclonal antibodies

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Preparation and crystallization of a human immunodeficiency virus p24-Fab complex. / Prongay, Andrew J.; Smith, Thomas; Rossmann, Michael G.; Ehrlich, Lorna S.; Carter, Carol A.; McClure, Jan.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 24, 1990, p. 9980-9984.

Research output: Contribution to journalArticle

Prongay, Andrew J. ; Smith, Thomas ; Rossmann, Michael G. ; Ehrlich, Lorna S. ; Carter, Carol A. ; McClure, Jan. / Preparation and crystallization of a human immunodeficiency virus p24-Fab complex. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 24. pp. 9980-9984.
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AU - McClure, Jan

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