Purification and characterization of the specific binding protein for platelet activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) from human platelets.

Nishihira, Jun; Ishibashi, Teruo; Imai, Yoh; Muramatsu, Toshiya

doi:10.1620/tjem.147.145

Cited by 18 publications

(6 citation statements)

References 19 publications

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“…39 kDa. Of course, this is in sharp contrast with the size of the solubilized receptor observed by gel filtration in our study, as well as in previous reports (Chau & Jii, 1988;Nishihara et al, 1985). The study by Honda et al (1991) showed no characterization of the expressed protein, so it is difficult to know whether the solubilized receptor may have a higher molecular mass than expected due to either detergent and lipid micelles, or due to other associated proteins that remain tightly bound.…”

Section: Discussioncontrasting

confidence: 99%

“…The solubilization of functionally active PAF receptors for the purpose of characterization and eventual purification has not previously been documented. Therefore this receptor system is poorly understood, with only a minimum of attention being given to the nature of the receptor's components (Nishihara et al, 1985;Chau & Jii, 1988;Chau et al, 1989). In this report we have described our attempts to systematically examine the ability of a series of detergents to solubilize the PAF receptor in a form that allows it to retain its binding activity.…”

Section: Discussionmentioning

confidence: 99%

“…2(b) shows the profile obtained from CHAPS solubilization ofplatelets pre-bound with [3H]PAF. Only 30 % of the radioactivity was recovered in a high-molecular mass fraction that presumably contained the PAF receptor complex (Nishihara et al, 1985;Chau & Jii, 1988). The remaining radioactivity migrated at the position expected for [3H]PAF alone, which associated with the BSA present in the column buffer ( Fig.…”

Section: Detergent Solubilization After Pre-binding Of I3hipafmentioning

confidence: 90%

“…Two reports have shown that membranes in which PAF was pre-bound to its receptor could be solubilized using detergents. Nishihara et al (1985) solubilized human platelet membranes pre-incubated with [3H]PAF using Triton X-100 and showed that the label was associated with a complex having a molecular mass of 160 kDa. Chau & Jii (1988) attempted a similar procedure with rabbit platelet membranes using digitonin, and showed that the label migrated with a molecular mass of 220 kDa.…”

Section: Introductionmentioning

confidence: 99%

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Solubilization of a functionally active platelet-activating factor receptor from rabbit platelets

Rogers

Duronio

Wong

et al. 1991

Biochemical Journal

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Binding of platelet-activating factor (PAF) to a specific high-affinity membrane receptor has been demonstrated in numerous cell types, but very little is known about the molecular nature of this receptor. The receptor from rabbit platelets was solubilized using CHAPS, digitonin, octyl glucoside, Nonidet P-40 or sodium cholate, either with pre-bound [3H]PAF or in the absence of ligand. We have been able to demonstrate for the first time that the receptor solubilized with CHAPS, in the absence of ligand, could retain its binding activity. It migrated as a high molecular mass complex (greater than 350 kDa) on a Bio-Gel A-0.5 m gel filtration column. Binding to solubilized receptor rapidly reached an equilibrium at room temperature, but was much slower at 0 degrees C. Scatchard plots were used to calculate the number (approx. 100 per cell) and the affinity (Kd 2.5 +/- 1.4 nM) of the solubilized receptors. These values were comparable with those obtained from whole-cell binding experiments. Competition by PAF antagonists also verified that the assay was measuring PAF receptor binding activity. The presence of a protein in the receptor complex was demonstrated by heat and trypsin inactivation of binding activity. Trypsin had no effect on binding of PAF to whole cells, but was able to decrease binding activity in solubilized receptor preparations. Attempts to demonstrate the involvement of a glycoprotein by use of various lectin columns proved unsuccessful. The latter results are consistent with findings suggesting that the binding site of the PAF receptor may not be exposed at the cell surface.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Detergent Solubilization After Pre-binding Of I3hipafmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

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Solubilization of a functionally active platelet-activating factor receptor from rabbit platelets

Rogers

Duronio

Wong

et al. 1991

Biochemical Journal

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“…Early attempts to purify and to characterize PAF receptor proteins from different sources, mainly from rabbit and human platelets [43][44][45] have been unsuccessful. Using photoaffinity labelling techniques, PAF binding proteins in rabbit platelet membranes have been characterized [46].…”

Section: Figure 3 Schematic Representation Of Paf Receptormentioning

confidence: 99%

Platelet-activating factor: receptors and signal transduction

Chao

Olson

1993

Biochemical Journal

432

270

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During the past two decades, studies describing the chemistry and biology of PAF have been extensive. This potent phosphoacylglycerol exhibits a wide variety of physiological and pathophysiological effects in various cells and tissues. PAF acts, through specific receptors and a variety of signal transduction systems, to elicit diverse biochemical responses. Several important future directions can be enumerated for the characterization of PAF receptors and their attendant signalling mechanisms. The recent cloning and sequence analysis of the gene for the PAF receptor will allow a number of important experimental approaches for characterizing the structure and analysing the function of the various domains of the receptor. Using molecular genetic and immunological technologies, questions relating to whether there is receptor heterogeneity, the precise mechanism(s) for the regulation of the PAF receptor, and the molecular details of the signalling mechanisms in which the PAF receptor is involved can be explored. Another area of major significance is the examination of the relationship between the signalling response(s) evoked by PAF binding to its receptor and signalling mechanisms activated by a myriad of other mediators, cytokines and growth factors. A very exciting recent development in which PAF receptors undoubtedly play a role is in the regulation of the function of various cellular adhesion molecules. Finally, there remain many incompletely characterized physiological and pathophysiological situations in which PAF and its receptor play a crucial signalling role. Our laboratory has been active in the elucidation of several tissue responses in which PAF exhibits major autocoid signalling responses, e.g. hepatic injury and inflammation, acute and chronic pancreatitis, and cerebral stimulation and/or trauma. As new experimental strategies are developed for characterizing the fine structure of the molecular mechanisms involved in tissue injury and inflammation, the essential role of PAF as a primary signalling molecule will be affirmed. Doubtless the next 20 years of experimental activity will be even more interesting and productive than the past two decades.

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Natural antagonists of platelet‐activating factor

Hosford

Mencia‐Huerta

Page

et al. 1988

Phytotherapy Research

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Several groups of antagonists of platelet-activating factor (PAF) are reviewed. The characteristics of PAF binding sites on mammalian cell membranes are described together with the activities of non-specific and synthetic PAF-antagonists. The specificity of the ginkgolides to bind to PAF-receptors is discussed and the effects of these natural products in a range of biological systems is emphasized. Other groups of natural PAF-antagonists described are the lignans and gliotoxin related compounds. The structural chemistry, structure-activity relationships and pharmacological properties of these drugs are compared to the ginkgolides.

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Purification and characterization of the specific binding protein for platelet activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) from human platelets.

Cited by 18 publications

References 19 publications

Solubilization of a functionally active platelet-activating factor receptor from rabbit platelets

Solubilization of a functionally active platelet-activating factor receptor from rabbit platelets

Platelet-activating factor: receptors and signal transduction

Natural antagonists of platelet‐activating factor

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