Nieto J, Veelaert D, Derua R, Waelkens E, Cerstiaens A, Coast G, Devreese B, Van Beeumen J, Calderon J, De Loof A, Schoofs L
Zoological Institute, Katholieke Universiteit Leuven, Naamsestraat 59, Leuven, B-3000, Belgium.
Biochem Biophys Res Commun. 1998 Jul 20;248(2):406-11. doi: 10.1006/bbrc.1998.8964.
This paper reports the purification of three myotropic neuropeptides from the white shrimp Penaeus vannamei. The central nervous systems of 3500 shrimps were extracted in an acidified solvent, after which four to five HPLC column systems were used to obtain pure peptides. A cockroach hindgut muscle contraction bioassay was used to monitor all collected fractions. The pure peptides were submitted to Edman degradation based automated microsequencing. Mass spectrometry and chemical synthesis confirmed the sequences. Ala-Pro-Ser-Gly-Phe-Leu-Gly-Met-Arg-NH2 (Pev-tachykinin, 934.1 Da) belongs to the tachykinin family with identified members in all vertebrate classes and some invertebrate classes: arthropods, annelids and molluscs. A very specific Pev-tachykinin antiserum was developed, which labels 4 neurosecretory cells in the brain. Ala-Ser-Phe-Ser-Pro-Trp-Gly-NH2 (Pev-kinin 1, 749.8 Da) and Asp-Phe-Ser-Ala-Trp-Ala-NH2 (Pev-kinin 2, 694.7 Da) are the first crustacean kinins. Pev-kinin 2 is the first kinin with a Trp-Ala-NH2 instead of a kinin-typical Trp-Gly-NH2 carboxyterminus.
本文报道了从凡纳滨对虾中纯化出三种促肌神经肽的过程。用酸化溶剂提取3500只虾的中枢神经系统,之后使用四到五个高效液相色谱柱系统来获取纯肽。采用蟑螂后肠肌肉收缩生物测定法监测所有收集的组分。将纯肽进行基于埃德曼降解的自动化微量测序。质谱分析和化学合成证实了其序列。丙氨酸-脯氨酸-丝氨酸-甘氨酸-苯丙氨酸-亮氨酸-甘氨酸-甲硫氨酸-精氨酸-氨基(凡纳滨对虾速激肽,934.1道尔顿)属于速激肽家族,在所有脊椎动物类群以及一些无脊椎动物类群(节肢动物、环节动物和软体动物)中都有已鉴定的成员。制备了一种非常特异的凡纳滨对虾速激肽抗血清,它能标记脑中的4个神经分泌细胞。丙氨酸-丝氨酸-苯丙氨酸-丝氨酸-脯氨酸-色氨酸-甘氨酸-氨基(凡纳滨对虾激肽1,749.8道尔顿)和天冬氨酸-苯丙氨酸-丝氨酸-丙氨酸-色氨酸-丙氨酸-氨基(凡纳滨对虾激肽2,694.7道尔顿)是首批甲壳类激肽。凡纳滨对虾激肽2是首个羧基末端为色氨酸-丙氨酸-氨基而非激肽典型的色氨酸-甘氨酸-氨基的激肽。