The pore size and permeability control of a glucose-responsive gating membrane with plasma-grafted poly (acrylic acid) (PAAC) gates and covalently bound glucose oxidase (GOD) enzymes were investigated systematically. The PAAC-grafted porous polyvinylidene fluoride (PVDF) membranes with a wide range of grafting yields were prepared using a plasma-graft pore-filling polymerization method, and the immobilization of GOD was carried out by a carbodiimide method. The linear grafted PAAC chains in the membrane pores acted as the pH-responsive gates or actuators. The immobilized GOD acted as the glucose sensor and catalyzer; it was sensitive to glucose and catalyzed the glucose conversion to gluconic acid. The experimental results showed that the glucose responsivity of the solute diffusional permeability through the proposed membranes was heavily dependent on the PAAC grafting yield, because the pH-responsive change of pore size governed the glucose-responsive diffusional permeability. It is very important to design a proper grafting yield for obtaining an ideal gating response. For the proposed gating membrane with a PAAC grafting yield of 1.55%, the insulin permeation coefficient after the glucose addition (0.2mol/l) was about 9.37 times that in the absence of glucose, presenting an exciting result on glucosesensitive self-regulated insulin permeation.