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J C Mani

Publications and source records attributed to J C Mani.

120 records · Page 7Linked to original sources
PubMedDate not supplied

Study of hydrophobic interactions between acylated proteins and phospholipid bilayers using BIACORE.

Intracellular proteins of eukaryotic cells are frequently covalently modified by the addition of long chain fatty acids. These modifications are thought to allow otherwise soluble proteins to associate with membranes by lipid-lipid based hydrophobic interactions. The purpose of this work was to quantify the effect of acyl chain length on hydrophobic interactions between acylated proteins and phospholipid monolayers. The binding of an artificially acylated model protein to electrically neutral phospholipids was studied by surface plasmon resonance, using BIACORE. Kinetic rates for the binding of bovine pancreatic ribonuclease A (RNase A), monoacylated on its N-terminal lysine with fatty acids of 10, 12, 14, 16 or 18 carbon atoms, to phospholipids on hydrophobic sensor chips, were measured. Unlike unmodified ribonuclease, acylated RNase A bound to the phospholipids, and the association level increased with the acyl chain length to reach a maximum for C16. Reproducible kinetics were obtained which did not fit a 1:1 Langmuir model but rather a two-step binding profile.

Acylation↗
PubMedDate not supplied

Lymphocyte membrane adenosine receptors coupled to adenylate cyclase: properties and occurrence in various lymphocyte subclasses.

Lymphocytes possess membrane adenosine receptors through which this nucleoside or analogs activate adenylate cyclase. Several adenosine analogs - 5'-N-ethylcarboxamide adenosine (NECA), N6-(L-2-phenyl-isopropyl) adenosine (PIA), 2-chloro adenosine (Cl-Ado) - were tested on the adenylate cyclase of mouse thymocyte homogenates; the order of potencies: NECA greater than Cl-Ado greater than PIA is consistent with the order which has been assumed to be characteristic of Ra type adenosine receptors. The stimulation of thymocyte adenylate cyclase by NECA was biphasic while the responses to Cl-Ado and PIA were not, which suggests the existence of Ra sub-types of adenosine receptors. Moreover, these adenosine receptors were found to be present in several lymphocyte subclasses: mature and immature mouse thymocytes, B and T mouse splenocytes, human peripheral blood lymphocytes, and also in lymphoblastoid cell lines. No crucial differences were observed between the adenylate cyclase stimulations of the various population by NECA.

Adenosine↗
PubMedDate not supplied

Epitope analysis of human IL-6 receptor gp80 molecule with monoclonal antibodies.

Gp80 human IL-6R was studied using 7 murine mAb (M37, M91, M113, M139, M164, M182 and M195) obtained after fusion of splenocytes of Balb/c mice immunised with a mixture of recombinant IL-6 receptor (rIL-6R) and cells from 2 cell lines expressing IL-6R. These were U266, which is IL-6 independent and XG-1 which is IL-6-dependent. In ELISA the 7 mAb reacted against the rIL-6R and against the natural soluble form found in plasma (nIL-6R), which both lack transmembrane and cytoplasmic domains. However, M195 reacted less with the natural than with the recombinant soluble IL-6R. Using FACS analysis, the 7 mAb were shown to bind to U266 cells but not to the Namalva cell line which is deprived of IL-6R. This showed that they all recognised the membrane form of the IL-6R. Three of the anti-IL-6R mAb reacted with rIL-6R by Western blotting. Four different epitopes of the molecule were identified, either by cross-blocking experiments of mAb binding to IL6R in ELISA or by the biosensor Biacore technology. A group of 4 mAb (M37, M113, M139 and M164) and another mAb (M195) identified 2 different epitopes involved in IL-6 binding. These antibodies were able to inhibit the binding of IL-6 to IL-6R and the proliferation of the IL-6-dependent XG-1 cell line. M91 and M182 recognized 2 other epitopes that were not involved in IL-6 binding. As expected, M91 did not inhibit XG-1 proliferation; in contrast, M182 interfered with the proliferative response of the XG-1 cell line.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗