Molecular cloning and analysis of in vivo expression of murine P- selectin

We, Sanders; Wilson, Raymond W.; Ballantyne, C M B; Beaudet, A. L.

doi:10.1182/blood.v80.3.795.bloodjournal803795

Cited by 25 publications

(30 citation statements)

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“…Residues in bold are numbered according to their positions in the human sequence. The entire sequence is highly conserved, and residues Ser772 through to Ala784 (underlined) are invariant, in human [34], porcine [35], bovine [36], murine [37], rat [38], canine and ovine sequences. The canine sequence has the GenBank™ accession number M88170 (Manning AM, Sanders WE Jr, Kukielka GL, Dore M, Rosenbloom CL, Hawkins HL, Michael LH, Entman ML, Smith CW, Beaudet AL, Anderson DC.…”

mentioning

confidence: 99%

A Third Specificity‐Determining Site in μ2 Adaptin for Sequences Upstream of YxxΦ Sorting Motifs

Owen

Setiadi

Evans

et al. 2001

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Internalization signals of the YxxF type (F =bulky hydrophobic side chain) interact with the m2 chain of AP-2 adaptors. Internalization activity is intolerant of non-conservative substitution of either the tyrosine or the F side chains, which bind to hydrophobic pockets in m2 adaptin in a conformation described as 'a two pinned plug into a socket'. P-selectin, a type I transmembrane protein, contains the YxxF-like sequence YGVF in its cytoplasmic domain, but substitution of either the tyrosine or phenylalanine with alanine in the full-length protein causes only small changes in the rate of endocytosis. It is shown here that the sequence YGVF contained within a peptide corresponding to the 17 COOH-terminal amino acids of P-selectin binds to m2 adaptin in the same fashion previously seen for other YxxF motifs. In addition, the P-selectin peptide binds to a third hydrophobic pocket in m2 adaptin through a leucine at position Y− 3 in the peptide. This structure suggests that some sequences can function as a 'three pinned plug', in which internalization activity is not critically dependent on any one of the three interacting side chains.

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mentioning

confidence: 99%

A Third Specificity‐Determining Site in μ2 Adaptin for Sequences Upstream of YxxΦ Sorting Motifs

Owen

Setiadi

Evans

et al. 2001

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“…Up‐regulation of endothelial P‐selectin in vivo has been demonstrated following complement‐induced injury [23]. Recent animal studies have suggested that P‐selectin expression can be induced by cytokines including IL‐1 and tumour necrosis factor (TNF) [24, 25]. This may represent one mechanism by which endogenous GC could regulate P‐selectin expression, given that ADX has been previously shown to increase IL‐1 release in in vivo and in vitro studies [26, 27] and to influence the response to IL‐1 and TNF [28].…”

Section: Discussionmentioning

confidence: 99%

Endogenous glucocorticoids modulate experimental anti-glomerular basement membrane glomerulonephritis

Leech

Huang

Morand

et al. 2000

Clinical and Experimental Immunology

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SUMMARYThe influence of endogenous glucocorticoids (GC) on glomerular injury was studied in a rat model of heterologous anti-glomerular basement membrane (GBM) glomerulonephritis (GN). Sprague-Dawley rats underwent adrenalectomy (ADX) or sham-operation 3 days prior to i.v. administration of both nephritogenic (100 mg/g) and subnephritogenic (50 mg/g) doses of sheep anti-rat GBM globulin. Administration of a subnephritogenic dose of anti-GBM globulin resulted in GN in adrenalectomized animals only. Similarly, ADX performed prior to administration of anti-GBM in the nephritogenic dose range resulted in exacerbation of GN compared with sham-operated animals (24 h protein excretion: 190·8 Ϯ 32·8 versus 42·5 Ϯ 2·6 mg/24 h; P < 0·005). In ADX animals receiving subnephritogenic doses of anti-GBM injury was manifested by abnormal proteinuria (62·7 Ϯ 5·8 mg/24 h), accumulation of neutrophils which peaked at 6 h (7·2 Ϯ 1·37 neutrophils per glomerular cross-section (neut/gcs)) and macrophage accumulation in glomeruli at 24 h (6·8 Ϯ 1·2 macrophages/gcs). Sham-adrenalectomized animals given the same dose of anti-GBM globulin developed minimal or no glomerular injury: urinary protein excretion (8·7 Ϯ 1·5 mg/24 h, P < 0·001); neutrophils (0·2 Ϯ 0·04 neutrophils/gcs, P < 0·001); macrophages (1·2 Ϯ 0·5 macrophages/gcs, P < 0·001). The increased cellular recruitment to glomeruli in adrenalectomized animals was associated with glomerular endothelial P-selectin expression. P-selectin expression was not detected in sham-operated rats after anti-GBM injection. Complement deposition in glomeruli was minimal in both groups. Physiologic GC replacement of ADX rats receiving subnephritogenic-dose anti-GBM reversed the observed susceptibility to GN development, with urinary protein excretion (7·8 Ϯ 1·12, P < 0·005) and no detectable P-selectin expression or leucocyte accumulation in glomeruli. These results suggest that endogenous GC modulate heterologous anti-GBM nephritis in rats and that this may be attributable, in part, to regulation of P-selectin expression.

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“…A certain number of amino acids are conserved, in a region spanning residues 359-457, in human and rat, but not in mouse P-selectin ( Figure 3). In that context it is of interest to note that mouse P-selectin is highly homologous, with the exception of SCR 2 [23], to its human counterpart. Homologue replacement mutagenesis of 10 conserved amino acids, spanning residues 359-457 was performed to locate those implicated in the LYP20 epitope.…”

Section: Homologue Replacement Mutagenesis Of the Region Spanning Resmentioning

confidence: 99%

Mapping the epitope of a functional P-selectin monoclonal antibody (LYP20) to a short complement-like repeat (SCR 4) domain: use of human–mouse chimaera and homologue-replacement mutagenesis

Ruchaud-Sparagano

Malaud

Gayet

et al. 1998

Biochemical Journal

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P-selectin (CD62P), an adhesion molecule localized in platelet alpha-granules and endothelial cell Weibel-Palade bodies, is rapidly expressed on the surface of activated cells. This adhesion molecule, a member of the selectin family, mediates leucocyte interactions with activated platelets or endothelial cells. The aim of this study was to identify and characterize the epitope of a functional blocking P-selectin monoclonal antibody (mAb), LYP20. LYP20 recognizes human or rat, but not mouse, P-selectin. Human/mouse chimaeras and wild-type constructs, modified by homologue replacement mutagenesis, were expressed in COS cells. Blocking anti-(P-selectin) mAbs (G1, G3 or CLB-thromb/6) were observed, by flow cytometry, to bind to the lectin-like domain. In contrast, LYP20 was found to bind to one of the P-selectin short complement-like repeats (SCR domain 4). Homologue replacement mutagenesis of SCR domain 4 (region delineated by amino acid residues 359-457) identified three amino acids (Cys412-->Ser, Cys416-->Ser or Arg415-->Lys) as being implicated in the LYP20 epitope. Deleting the region bearing the LYP20 epitope, from a wild-type CD62P construct, showed a decrease in polymorphonuclear leucocyte (PMNL) binding to transfected COS cells. In addition, mutation of one of the three amino acids, implicated in the LYP20 epitope, markedly affected PMNL binding to transfected COS cells but did not affect the binding of mAbs G1 and CLB-thromb/6. These results are the first to indicate (1) that a functional blocking anti-P-selectin mAb binds to SCR 4, a site other than the lectin-like/epidermal growth factor-like domain, and (2) that SCR domain 4 has a functional role in P-selectin-leucocyte interactions.

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Molecular cloning and analysis of in vivo expression of murine P- selectin

Cited by 25 publications

References 0 publications

A Third Specificity‐Determining Site in μ2 Adaptin for Sequences Upstream of YxxΦ Sorting Motifs

A Third Specificity‐Determining Site in μ2 Adaptin for Sequences Upstream of YxxΦ Sorting Motifs

Endogenous glucocorticoids modulate experimental anti-glomerular basement membrane glomerulonephritis

Mapping the epitope of a functional P-selectin monoclonal antibody (LYP20) to a short complement-like repeat (SCR 4) domain: use of human–mouse chimaera and homologue-replacement mutagenesis

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