Mutations inside but not outside the peptide binding cleft of the H-2 Ld molecule affects CTL recognition and binding of the nucleoprotein peptide from the lymphocytic chroriomeningitis

Catarina E. Hioe, Denise M. McKinney, Jeffrey A Frelinger, Minnie McMillan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

In order to investigate the role of residues inside and outside the peptide binding cleft of the L2 molecule in peptide presentation to cytotoxic T lymphocytes (CTL), we constructed a series of point mutations in the Ld gene. We determined the effects of the mutations in the Ld molecule on the binding and recognition of an Ld-restricted CTL epitope derived from the nucleoprotein (NP) of the lymphocytic phoriomeningitis virus (LCMV). Each of the mutations within the Ld peptide binding cleft resulted in a complete loss of CTL recognition. Addition of the LCMV NP peptide to cells expressing these mutants did not increase surface Ld expression, suggesting that the mutations altered peptide binding. Mutations involving pockets D and E within the cleft affected LCMV peptide binding and recognition as drastically as those in pocket B, which was predicted to interact with a main anchor residue of the peptide. In striking contrast, the mutations located outside the cleft did not change either recognition or binding. These results demonstrate that the Ld residues in the peptide binding cleft are the main determinants dictating LCMV NP peptide binding, and that the residues in each of the pockets within the cleft play a role in this interaction. Surprisingly, one mutation outside the peptide binding cleft, T92S, abrogated CTL lysis of target cells treated with the LCMV NP peptide, but not virus-infected cells. These data show that this mutation selectively altered the presentation of the LCMV NP peptide introduced to the cell exogenously, but not endogenously. This implies that the pathway by which peptides associate with class I molecules within the cell differs from that of exogenous peptide binding.

Original languageEnglish (US)
Pages (from-to)222-229
Number of pages8
JournalImmunogenetics
Volume40
Issue number3
DOIs
StatePublished - Jul 1994
Externally publishedYes

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Nucleoproteins
Cytotoxic T-Lymphocytes
Peptides
Mutation
Viruses
Virus Attachment
T-Lymphocyte Epitopes
Point Mutation

ASJC Scopus subject areas

  • Immunology
  • Genetics

Cite this

Mutations inside but not outside the peptide binding cleft of the H-2 Ld molecule affects CTL recognition and binding of the nucleoprotein peptide from the lymphocytic chroriomeningitis. / Hioe, Catarina E.; McKinney, Denise M.; Frelinger, Jeffrey A; McMillan, Minnie.

In: Immunogenetics, Vol. 40, No. 3, 07.1994, p. 222-229.

Research output: Contribution to journalArticle

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abstract = "In order to investigate the role of residues inside and outside the peptide binding cleft of the L2 molecule in peptide presentation to cytotoxic T lymphocytes (CTL), we constructed a series of point mutations in the Ld gene. We determined the effects of the mutations in the Ld molecule on the binding and recognition of an Ld-restricted CTL epitope derived from the nucleoprotein (NP) of the lymphocytic phoriomeningitis virus (LCMV). Each of the mutations within the Ld peptide binding cleft resulted in a complete loss of CTL recognition. Addition of the LCMV NP peptide to cells expressing these mutants did not increase surface Ld expression, suggesting that the mutations altered peptide binding. Mutations involving pockets D and E within the cleft affected LCMV peptide binding and recognition as drastically as those in pocket B, which was predicted to interact with a main anchor residue of the peptide. In striking contrast, the mutations located outside the cleft did not change either recognition or binding. These results demonstrate that the Ld residues in the peptide binding cleft are the main determinants dictating LCMV NP peptide binding, and that the residues in each of the pockets within the cleft play a role in this interaction. Surprisingly, one mutation outside the peptide binding cleft, T92S, abrogated CTL lysis of target cells treated with the LCMV NP peptide, but not virus-infected cells. These data show that this mutation selectively altered the presentation of the LCMV NP peptide introduced to the cell exogenously, but not endogenously. This implies that the pathway by which peptides associate with class I molecules within the cell differs from that of exogenous peptide binding.",
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