Binding of protein chemotactic factors to the surfaces of neutrophil leukocytes and its modification with lipid-specific bacterial toxins.
The binding to neutrophil leukoyctes of human serum albumin (HSA), which is chemokinetic  for leukocytes, i.e. influences their rate of locomotion, and of alkali-denatured HSA, which is chemotactic for leukocytes, i.e. influences their direction of locomotion, was studied. Native serum albumin showed low affinity binding to the neutrophil surface. Denatured serum albumin showed saturable binding with a Ka of approximately 1-(6) litres per mole to about 10(6) binding sites per cell. Another protein chemotactic factor, alpha5-casein, gave similar binding. These results exclude that chemotactic reactions to denatured proteins are mediated in a completely non-specific manner and suggest the presence on the cell of a restricted number of defined recognition sites. Binding was reduced following treatment of the cells with either of two lipid-specific bacterial toxins, perfringolysin, the theta-toxin of Clostridium perfringens, an oxygen-labile cholesterol-specific toxin, and Staphylococcus aureus Sphingomyelinase C. Both have previously been shown to reduce chemotactic reactions and both were used at doses which did not reduce cell viability. These results suggest an important, and possiblly direct, role for membrane lipid in the binding sites for chemotactic factors. Visual analysis of the behaviour of perfringolysin-treated neutrophils showed that these cells were still capable of chemotactic locomotion. The cells appeared to be less efficient than normal in detecting chemotactic gradients only when at a distance from the gradient source, a finding which is consistent with reduced binding of the chemotactic factor to the cell surface.
D001427:Bacterial Toxins / Q000378:metabolism*; D002633:Chemotaxis; D006026:Glycoside Hydrolases; D006801:Humans; D009000:Monocytes / Q000502:physiology; D009504:Neutrophils / Q000502:physiology*; D010447:Peptide Hydrolases; D010740:Phospholipases; D011485:Protein Binding; D012709:Serum Albumin; D013108:Sphingomyelin Phosphodiesterase
D001427:Bacterial Toxins; D012709:Serum Albumin; D010740:Phospholipases; D013108:Sphingomyelin Phosphodiesterase; D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases