Production and crystallization of MHC class I B allele single peptide complexes

Reid, S. W. J.; Smith, Kathrine J.; Jakobsen, Bent K.; O’Callaghan, C A; Reyburn, Hugh; Harlos, K.; Stuart, David I.; McMichael, Andrew J.; Bell, John I.; Jones, E Y

doi:10.1016/0014-5793(96)00226-8

Cited by 36 publications

(34 citation statements)

References 36 publications

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“…2). Similar to the other LRC receptors, the first ␤-strand (A strand) in D1 of LILRA5 is split into two shorter strands (A and AЈ) by a cis-proline (Pro 12 ) and shared between the two ␤-strands (B and G strands). However, residues 106 -116 of D2, which corresponds to the AЈ strand of D1 in the other LRC receptors, was disordered.…”

Section: Resultsmentioning

confidence: 99%

“…For introduction of the restriction enzyme sites (NdeI and HindIII), 5Ј-GGA-ATTCCATATGGGGAACCTCTCCAAAGCCAC-3Ј (forward) and 5Ј-TAGGCAAGCTTATGAGACCGGAATCTCCAG-GAG-3Ј (reverse) primers were used. The resultant PCR fragment including the whole extracellular domain (residues with additional N-terminal methionine was digested with the restriction enzymes NdeI and HindIII and ligated into the pGMT7 expression vector (12). Inclusion bodies of LILRA5 were expressed in Escherichia coli strain BL21(DE3)pLysS and prepared as previously reported for LILRB1 and LILRB2 (7).…”

Section: Methodsmentioning

confidence: 99%

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Crystal Structure of the Human Monocyte-activating Receptor, “Group 2” Leukocyte Ig-like Receptor A5 (LILRA5/LIR9/ILT11)

Shiroishi

Kajikawa

Kuroki

et al. 2006

Journal of Biological Chemistry

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Human leukocyte Ig-like receptor B1 (LILRB1) and B2 (LILRB2) belong to "Group 1" receptors and recognize a broad range of major histocompatibility complex class I molecules (MHCIs). In contrast, "Group 2" receptors show low similarity with LILRB1/B2, and their ligands remain to be identified. To date, the structural and functional characteristics of Group 2 LILRs are poorly understood. Here we report the crystal structure of the extracellular domain of LILRA5, which is an activating Group 2 LILR expressed on monocytes and neutrophils. Unexpectedly, the structure showed large changes in structural conformation and charge distribution in the region corresponding to the (1). The LILR family has two or four tandem Ig-like extracellular domains and is divided into three functional categories: inhibitory, activating, and secreted receptors (2). The inhibitory LILR receptors have long cytoplasmic tails with two to four immunoreceptor tyrosine-based inhibitory motifs, in which the phosphorylation of tyrosine residues recruits the Src homology 2 domain-containing tyrosine phosphatase-1, transducing negative signals to inhibit activation signals. In contrast, the activating receptors have short cytoplasmic tails lacking immunoreceptor tyrosine-based inhibitory motifs and an arginine residue in the transmembrane region, which binds to negatively charged residues of FcR␥ harboring the immunoreceptor tyrosine-based activation motif. The LILRs are mainly expressed on myelomonocytic cells, although some are also expressed on a wide range of leukocytes. LILRA5 (LIR9/ILT11/CD85f) is expressed as both a membrane-bound activating and a secreted form (3). The membrane-bound form of LILRA5 has two Ig-like domains in the extracellular region with typical features of activating receptors (an arginine in the transmenbrane region and a short cytoplasmic domain), whereas the secreted form only has the extracellular domain. In contrast with LILRB1, which is expressed on a range of leukocytes, the expression of the activating form of LILRA5 has been detected on CD14 ϩ monocytes (3). The expression of LILRA5 has also been detected in neutrophils and monocytes but only at the mRNA level (3). Cross-linking of LILRA5 molecules on monocytes using anti-LILRA5 antibody induced the mobilization of calcium and the secretion of cytokines released in the early stages of inflammatory responses, such as interleukin-

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Crystal Structure of the Human Monocyte-activating Receptor, “Group 2” Leukocyte Ig-like Receptor A5 (LILRA5/LIR9/ILT11)

Shiroishi

Kajikawa

Kuroki

et al. 2006

Journal of Biological Chemistry

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“…The ectodomain of KIR2DL1 (residues 1-224) and CD8␣ homodimer (CD8␣␣) (residues 1-120) were produced as described (27,29). ILT2D1D2 and ILT4D1D2 with or without a fixed amount of KIR2DL1(38 M), which was almost saturated, were flowed over the immobilized HLACw*0401.…”

Section: Surface Plasmon Resonance (Spr)mentioning

confidence: 99%

Human inhibitory receptors Ig-like transcript 2 (ILT2) and ILT4 compete with CD8 for MHC class I binding and bind preferentially to HLA-G

Shiroishi

Tsumoto

Amano

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

477

443

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Ig-like transcript 4 (ILT4) (also known as leukocyte Ig-like receptor 2, CD85d, and LILRB2) is a cell surface receptor expressed mainly on myelomonocytic cells, whereas ILT2 (also known as leukocyte Ig-like receptor 1, CD85j, and LILRB1) is expressed on a wider range of immune cells including subsets of natural killer and T cells. Both ILTs contain immunoreceptor tyrosine-based inhibitory receptor motifs in their cytoplasmic tails that inhibit cellular responses by recruiting phosphatases such as SHP-1 (Src homology 2 domain containing tyrosine phosphatase 1). Although these ILTs have been shown to recognize a broad range of classical and nonclassical human MHC class I molecules (MHCIs), their precise binding properties remain controversial. We have used surface plasmon resonance to analyze the interaction of soluble forms of ILT4 and ILT2 with several MHCIs. Although the range of affinities measured was quite broad (K d ‫؍‬ 2-45 M), some interesting differences were observed. ILT2 generally bound with a 2-to 3-fold higher affinity than ILT4 to the same MHCI. Furthermore, ILT2 and ILT4 bound to HLA-G with a 3-to 4-fold higher affinity than to classical MHCIs, suggesting that ILT͞HLA-G recognition may play a dominant role in the regulation of natural killer, T, and myelomonocytic cell activation. Finally, we show that ILT2 and ILT4 effectively compete with CD8 for MHCI binding, raising the possibility that ILT2 modulates CD8 ؉ T cell activation by blocking the CD8 binding as well as by recruiting inhibitory molecules through its immunoreceptor tyrosine-based inhibitory receptor motif.leukocyte Ig-like receptors ͉ major histocompatibility complex ͉ surface plasmon resonance ͉ natural killer cell ͉ coreceptor I g-like transcripts (ILTs) (also called leukocyte Ig-like receptors, CD85, or LILRB) are encoded by a family of immunoreceptor genes located at human chromosome 19q13.4. This locus is called the leukocyte receptor complex and includes, in addition to ILT genes, the genes encoding killer cell Ig-like receptors (KIRs), leukocyte-associated Ig-like receptors, NKp46, and the Fc␣ receptor (1). Although ILT2 is broadly expressed on monocytes, B cells, dendritic cells, and subsets of natural killer (NK) and T cells, ILT4 expression is largely confined to the myelomonocytic lineage (2-8). Both ILT2 and ILT4 have four tandem Ig-like extracellular domains and four and three immunoreceptor tyrosine-based inhibitory receptor motifs, respectively, in their cytoplasmic tails. Immunoreceptor tyrosine-based inhibitory receptor motifs recruit the protein tyrosine phosphatase SHP-1 (Src homology 2 domain containing phosphatase 1), which is thought to inhibit early signaling events triggered by stimulatory receptors. Indeed engagement of ILT2 on T cells has been shown to inhibit T cell antigen receptor (TCR) signaling and downstream events such as actin reorganization (9). Studies on CD8 ϩ cells suggest that ILT2 is expressed early on in contrast to KIRs, which are expressed primarily on the subset of stimulated CD8 ϩ cells tha...

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“…The major histocompatibility complex (MHC) molecule HLA B8 complexed with various peptides is being studied by Jones and co-workers from Oxford University Biophysics Department (Reid et al, 1996). Recognition by T cell receptors of MHC Class I molecules complexed with peptides from pathogens serves to trigger the cellular immune response.…”

Section: Beamline Id-2 Demonstration Experimentsmentioning

confidence: 99%

“…Recognition by T cell receptors of MHC Class I molecules complexed with peptides from pathogens serves to trigger the cellular immune response. Crystals of these complexes are of space group P21212l with unit-cell dimensions a --52, b = 81, c = 112 & (slight variations occur with cryocooling) (Reid et al, 1996). Although the complex crystals are well ordered, they are frequently small (0.1 × 0.05 x 0.2 ram) and necessitate high-intensity synchrotron radiation for high-resolution data.…”

Section: Beamline Id-2 Demonstration Experimentsmentioning

confidence: 99%

Calibration and Application of an X-ray Image Intensifier/ Charge-Coupled Device Detector for Monochromatic Macromolecular Crystallography

Hammersley

Brown²,

Burmeister³

et al. 1997

J Synchrotron Radiat

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Charge-coupled device (CCD)-based X-ray detectors allow data to be collected much more quickly (--,10 times) than with current on-line imaging-plate systems. At the ESRF, X-ray image intensifier/CCD detector systems have been developed. These have great potential as fast read-out detectors for macromolecular and other forms of crystallography. They are relatively large sensitive X-ray detectors but have two inherent weaknesses: convex detection surfaces leading to spatial distortion and non-uniformity of intensity response, and susceptibility to small changes in magnetic fields. A large improvement has been made to the accuracy obtained by non-uniformity of response calibration and correction, using fluorescence from doped lithium borate glasses. Monochromatic macromolecular crystallography demonstration experiments with external user groups have shown that high-quality results may be obtained under real experimental conditions.

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Production and crystallization of MHC class I B allele single peptide complexes

Cited by 36 publications

References 36 publications

Crystal Structure of the Human Monocyte-activating Receptor, “Group 2” Leukocyte Ig-like Receptor A5 (LILRA5/LIR9/ILT11)

Crystal Structure of the Human Monocyte-activating Receptor, “Group 2” Leukocyte Ig-like Receptor A5 (LILRA5/LIR9/ILT11)

Human inhibitory receptors Ig-like transcript 2 (ILT2) and ILT4 compete with CD8 for MHC class I binding and bind preferentially to HLA-G

Calibration and Application of an X-ray Image Intensifier/ Charge-Coupled Device Detector for Monochromatic Macromolecular Crystallography

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