Gibbons B H, Ogawa K, Gibbons I R
J Cell Biol. 1976 Dec;71(3):823-31. doi: 10.1083/jcb.71.3.823.
Rabbit antiserum prepared against an ATPase-containing tryptic fragment of dynein by Ogawa and Mohri (J. Biol. Chem. 250: 6476-6483) specifically inhibited the ATPase activity of dynein 1 and not that of dynein 2. Varying amounts of this antidynein 1 serum were added to demembranated sperm while they were swimming in reactivating solution containing 1 mM ATP. The sperm continued to form regularly propagated flagellar bending waves, but the beat frequency decreased gradually with time, the greater part of the change occurring in the first 15 min. The beat frequency after 1 h was a function of the amount of antiserum used, and could be as low as 1 Hz. The waveforms of the treated sperm resembled those of normal reactivated sperm except that the bend angles of both the principal and reverse bends were larger in the proximal portion of flagellum. The ATPase activity and corresponding beat frequency of sperm which had been pretreated with varying amounts of antidynein 1 serum for 15 min at 0 degrees C and then diluted were both decreased as a function of the amount of antiserum added, the ATPase activity of homogenized, nonmotile sperm also decreased upon pretreatment with antiserum, but the percentage decrease was less than for motile sperm. For moderate to low concentrations of antiserum, the rates of reaction with motile and with rigor sperm were almost identical. The overall results suggest that antidynein 1 inhibits the functioning of the dynein arms, probably by blocking the ATPase sites of the dynein 1.
小川和森(《生物化学杂志》250: 6476 - 6483)制备的针对动力蛋白含ATP酶胰蛋白酶片段的兔抗血清,特异性抑制动力蛋白1的ATP酶活性,而不抑制动力蛋白2的活性。在去膜精子于含1 mM ATP的再激活溶液中游动时,加入不同量的这种抗动力蛋白1血清。精子继续形成规则传播的鞭毛弯曲波,但搏动频率随时间逐渐降低,大部分变化发生在最初15分钟内。1小时后的搏动频率是所用抗血清量的函数,可低至1赫兹。处理后的精子波形与正常再激活精子的波形相似,只是在鞭毛近端,主弯曲和反向弯曲的弯曲角度都更大。在0℃用不同量的抗动力蛋白1血清预处理15分钟然后稀释的精子,其ATP酶活性和相应的搏动频率均随抗血清添加量的增加而降低,用抗血清预处理后,匀浆的、不活动精子的ATP酶活性也降低,但降低百分比低于活动精子。对于中低浓度的抗血清,其与活动精子和僵直精子的反应速率几乎相同。总体结果表明,抗动力蛋白1可能通过阻断动力蛋白1的ATP酶位点来抑制动力蛋白臂的功能。