Zhao Q, Sannier F, Piot J M
Laboratoire de Génie proteíque et cellulaire, Pôle Sciences et Technologies, La Rochelle, France.
Biochim Biophys Acta. 1996 Jun 7;1295(1):73-80. doi: 10.1016/0167-4838(96)00025-8.
The kinetics of appearance of hemorphins during peptic hydrolysis of bovine hemoglobin was investigated by reverse-phase high-performance liquid chromatography (RP-HPLC) coupled with a photodiode array detector. The degree of hydrolysis (DH) of hemoglobin by pepsin was determined and different defined DH of hydrolysates were obtained. The analysis of these hydrolysates by HPLC coupled with a photodiode array detector allowed us to identify and quantify the hemorphins in every hydrolysate and to determine the quantitative evolution of hemorphins as a function of DH. It indicated that hemoglobin was a direct precursor of LVV-hemorphin-5 and LVV-hemorphin-7. These peptides were demonstrated to be secondary substrates for pepsin to generate VV-hemorphin-5 and VV-hemorphin-7. Moreover, LVV-hemorphin-7 was more stable towards pepsin than LVV-hemorphin-5. The affinity of pepsin towards some peptidic bonds was also demonstrated.
采用反相高效液相色谱（RP-HPLC）结合光电二极管阵列检测器，研究了牛血红蛋白在胃蛋白酶消化水解过程中血啡肽的出现动力学。测定了胃蛋白酶对血红蛋白的水解度（DH），并获得了不同水解度的水解产物。通过HPLC结合光电二极管阵列检测器对这些水解产物进行分析，使我们能够鉴定和定量每种水解产物中的血啡肽，并确定血啡肽随水解度的定量变化。结果表明，血红蛋白是LVV-血啡肽-5和LVV-血啡肽-7的直接前体。这些肽被证明是胃蛋白酶产生VV-血啡肽-5和VV-血啡肽-7的二级底物。此外，LVV-血啡肽-7对胃蛋白酶的稳定性高于LVV-血啡肽-5。还证明了胃蛋白酶对某些肽键的亲和力。
