%0 Journal Article %1 Klotz1994c %A Klotz, K. N. %A Krotec, K. L. %A Gripentrog, J. %A Jesaitis, A. J. %D 1994 %J J Immunol %K & Acid Actins/metabolism Amino Cell Cell-Free Cytoskeleton/metabolism Data Formyl Humans Immunologic/*metabolism Leucyl-Phenylalanine/analogs Membrane/*ultrastructure Microfilaments/metabolism Molecular N-Formylmethionine Neutrophils/*metabolism/ultrastructure Peptide Peptide/*metabolism Sequence Solubility System derivatives/metabolism Receptor %N 2 %P 801-10 %T Regulatory interaction of N-formyl peptide chemoattractant receptors with the membrane skeleton in human neutrophils %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8283053 %V 152 %X The cytoskeleton and/or membrane skeleton has been implicated in the regulation of N-formyl peptide receptors. The coupling of these chemotactic receptors to the membrane skeleton was investigated in plasma membranes from unstimulated and desensitized human neutrophils using the photoreactive agonist N-formyl-met-leu-phe-lys-N epsilon-125I2(p-azidosalicylamido)ethyl-1,3'- dithiopropionate (fMLFK-125IASD). When membranes of unstimulated cells were solubilized in Triton-X 100, a detergent that does not disrupt actin filaments, only 50% of the photoaffinity-labeled receptors were solubilized sedimenting in sucrose density gradients at a rate consistent with previous reports. The remainder were found in the pellet fraction along with the membrane skeletal actin. Solubilization of the membranes in the presence of p-chloromercuriphenylsulfonic acid, elevated concentrations of KCl, or deoxyribonuclease I released receptors in parallel with actin. When membranes from neutrophils, desensitized by incubation with fMLFK-125IASD at 15 degrees C, were solubilized, nearly all receptors were recovered in the pellet fraction. Incubation of cells with the ligand at 4 degrees C inhibited desensitization partially and prevented the conversion of a significant fraction of receptors to the form associated with the membrane skeletal pellet. In these separations the photoaffinity-labeled receptors not sedimenting to the pellet cosedimented with actin. Approximately 25% of these receptors could be immunosedimented with antiactin antibodies suggesting that N-formyl peptide receptors may interact directly with actin. These results are consistent with a regulatory role for the interaction of chemotactic N-formyl peptide receptors with actin of the membrane skeleton.

@article{Klotz1994c, abstract = {The cytoskeleton and/or membrane skeleton has been implicated in the regulation of N-formyl peptide receptors. The coupling of these chemotactic receptors to the membrane skeleton was investigated in plasma membranes from unstimulated and desensitized human neutrophils using the photoreactive agonist N-formyl-met-leu-phe-lys-N epsilon-[125I]2(p-azidosalicylamido)ethyl-1,3'- dithiopropionate (fMLFK-[125I]ASD). When membranes of unstimulated cells were solubilized in Triton-X 100, a detergent that does not disrupt actin filaments, only 50% of the photoaffinity-labeled receptors were solubilized sedimenting in sucrose density gradients at a rate consistent with previous reports. The remainder were found in the pellet fraction along with the membrane skeletal actin. Solubilization of the membranes in the presence of p-chloromercuriphenylsulfonic acid, elevated concentrations of KCl, or deoxyribonuclease I released receptors in parallel with actin. When membranes from neutrophils, desensitized by incubation with fMLFK-[125I]ASD at 15 degrees C, were solubilized, nearly all receptors were recovered in the pellet fraction. Incubation of cells with the ligand at 4 degrees C inhibited desensitization partially and prevented the conversion of a significant fraction of receptors to the form associated with the membrane skeletal pellet. In these separations the photoaffinity-labeled receptors not sedimenting to the pellet cosedimented with actin. Approximately 25% of these receptors could be immunosedimented with antiactin antibodies suggesting that N-formyl peptide receptors may interact directly with actin. These results are consistent with a regulatory role for the interaction of chemotactic N-formyl peptide receptors with actin of the membrane skeleton.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Klotz, K. N. and Krotec, K. L. and Gripentrog, J. and Jesaitis, A. J.}, biburl = {https://www.bibsonomy.org/bibtex/2722085283dff1f542b3ec7af476de0bc/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {4b3701214105dd59b99c5319dab448a2}, intrahash = {722085283dff1f542b3ec7af476de0bc}, issn = {0022-1767 (Print) 0022-1767 (Linking)}, journal = {J Immunol}, keywords = {& Acid Actins/metabolism Amino Cell Cell-Free Cytoskeleton/metabolism Data Formyl Humans Immunologic/*metabolism Leucyl-Phenylalanine/analogs Membrane/*ultrastructure Microfilaments/metabolism Molecular N-Formylmethionine Neutrophils/*metabolism/ultrastructure Peptide Peptide/*metabolism Sequence Solubility System derivatives/metabolism Receptor}, month = {Jan 15}, note = {Klotz, K N Krotec, K L Gripentrog, J Jesaitis, A J R01-AI-22735/AI/NIAID NIH HHS/United States In Vitro Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states Journal of immunology (Baltimore, Md. : 1950) J Immunol. 1994 Jan 15;152(2):801-10.}, number = 2, pages = {801-10}, shorttitle = {Regulatory interaction of N-formyl peptide chemoattractant receptors with the membrane skeleton in human neutrophils}, timestamp = {2010-12-14T18:20:05.000+0100}, title = {Regulatory interaction of N-formyl peptide chemoattractant receptors with the membrane skeleton in human neutrophils}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8283053}, volume = 152, year = 1994 }