Alpha-casein-binding proteins of guinea pig macrophage membranes and their possible roles in chemotaxis.
The existence of specific binding proteins for alpha-casein, a well-known potent chemoattractant, on guinea pig peritoneal macrophages was demonstrated. Binding of 3H-alpha-casein to macrophages was found to be specific and reversible. Its association constant and the number of binding sites were 5.0 X 10(-6) M and 7.5 X 10(5) per cell, respectively. The presence of formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe), another chemoattractant, at a chemotactically active concentration (10(-11) to 10(-6) M) reduced this binding significantly. Pretreatment of macrophages with proteases such as trypsin and chymotrypsin decreased the number of binding sites for alpha-casein and the chemotactic response to alpha-casein. alpha-Casein binding proteins were isolated from radiolabeled macrophage surface membranes by affinity chromatography on a column of alpha-casein Sepharose 4B. SDS-polyacrylamide gel electrophoresis of the isolated proteins revealed three radioactive bands; two (MW 130,000 and 65,000) were glycoproteins, and the other (MW 100,000) was a protein which contained little carbohydrate. These three proteins were also isolated from the same source by affinity chromatography using a column of fMet-Leu-Phe-AH-Sepharose. This result, together with the observation that fMet-Leu-Phe inhibited the binding of alpha-casein to macrophages, indicated that alpha-casein and the tripeptide have at least partly common binding sites on guinea pig macrophages. Some of the proteins obtained in this work may constitute chemotactic receptors on the macrophage membrane.
D000818:Animals; D001894:Borohydrides; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002364:Caseins / Q000378:metabolism*; D002633:Chemotaxis; D004912:Erythrocytes / Q000502:physiology; D005260:Female; D006168:Guinea Pigs; D007106:Immune Sera; D007700:Kinetics; D008214:Lymphocytes / Q000502:physiology; D008264:Macrophages / Q000378:metabolism*; D009439:Neuraminidase / Q000494:pharmacology; D010084:Oxidation-Reduction; D010447:Peptide Hydrolases / Q000494:pharmacology; D011485:Protein Binding
D001894:Borohydrides; D002352:Carrier Proteins; D002364:Caseins; D007106:Immune Sera; C026046:alpha-casein-binding proteins; D009439:Neuraminidase; D010447:Peptide Hydrolases