PDF(Pubmed)
Abstract：
UNASSIGNED: Haemaphysalis flava is a notorious parasite for humans and animals worldwide. The organs of H. flava are bathed in hemolymph, which is a freely circulating fluid. Nutrients, immune factors, and waste can be transported to any part of the body via hemolymph. The main soluble components in hemolymph are proteins. However, knowledge of the H. flava proteome is limited.
UNASSIGNED: The hemolymph was collected from fully engorged H. flava ticks by leg amputation. Hemolymph proteins were examined by both blue native polyacrylamide gel electrophoresis (BN-PAGE) and sodium dodecyl sulfate PAGE (SDS-PAGE). Proteins extracted from the gels were further identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
UNASSIGNED: Two bands (380 and 520 kDa) were separated from tick hemolymph by BN-PAGE and were further separated into four bands (105, 120, 130, and 360 kDa) by SDS-PAGE. LC-MS/MS revealed that seven tick proteins and 13 host proteins were present in the four bands. These tick proteins mainly belonged to the vitellogenin (Vg) family and the α-macroglobulin family members. In silico structural analysis showed that these Vg family members all had common conserved domains, including the N-terminus lipid binding domain (LPD-N), the C-terminus von Willebrand type D domain (vWD), and the domain of unknown function (DUF). Additionally, two of the Vg family proteins were determined to belong to the carrier protein (CP) by analyzing the unique N-terminal amino acid sequences and the cleaving sites.
UNASSIGNED: These findings suggest that the Vg family proteins and α-macroglobulin are the primary constituents of the hemolymph in the form of protein complexes. Our results provide a valuable resource for further functional investigations of H. flava hemolymph effectors and may be useful in tick management.
UNASSIGNED: The hemolymph was collected from fully engorged H. flava ticks by leg amputation. Hemolymph proteins were examined by both blue native polyacrylamide gel electrophoresis (BN-PAGE) and sodium dodecyl sulfate PAGE (SDS-PAGE). Proteins extracted from the gels were further identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
UNASSIGNED: Two bands (380 and 520 kDa) were separated from tick hemolymph by BN-PAGE and were further separated into four bands (105, 120, 130, and 360 kDa) by SDS-PAGE. LC-MS/MS revealed that seven tick proteins and 13 host proteins were present in the four bands. These tick proteins mainly belonged to the vitellogenin (Vg) family and the α-macroglobulin family members. In silico structural analysis showed that these Vg family members all had common conserved domains, including the N-terminus lipid binding domain (LPD-N), the C-terminus von Willebrand type D domain (vWD), and the domain of unknown function (DUF). Additionally, two of the Vg family proteins were determined to belong to the carrier protein (CP) by analyzing the unique N-terminal amino acid sequences and the cleaving sites.
UNASSIGNED: These findings suggest that the Vg family proteins and α-macroglobulin are the primary constituents of the hemolymph in the form of protein complexes. Our results provide a valuable resource for further functional investigations of H. flava hemolymph effectors and may be useful in tick management.
摘要：
■黄藻是全世界人类和动物臭名昭著的寄生虫。黄曲霉的器官沐浴在血淋巴中，这是一种自由循环的流体。营养素,免疫因子，废物可以通过血淋巴输送到身体的任何部位。血淋巴中的主要可溶性成分是蛋白质。然而,H.flava蛋白质组的知识是有限的。
■通过截肢从完全充血的H.flava蜱中收集血淋巴。通过蓝色天然聚丙烯酰胺凝胶电泳(BN-PAGE)和十二烷基硫酸钠PAGE(SDS-PAGE)检查血淋巴蛋白。通过液相色谱-串联质谱(LC-MS/MS)进一步鉴定从凝胶中提取的蛋白质。
■通过BN-PAGE从蜱血淋巴中分离出两条带(380和520kDa),并通过SDS-PAGE进一步分离成四条带(105、120、130和360kDa)。LC-MS/MS显示在4个条带中存在7个蜱蛋白和13个宿主蛋白。这些蜱蛋白主要属于卵黄蛋白原(Vg)家族和α-巨球蛋白家族成员。计算机结构分析表明，这些Vg家族成员都具有共同的保守结构域，包括N末端脂质结合域（LPD-N），C端vonWillebrandD型结构域(vWD)，和未知函数域(DUF)。此外,通过分析独特的N-末端氨基酸序列和切割位点,确定Vg家族蛋白中的两种属于载体蛋白(CP)。
■这些发现表明，Vg家族蛋白和α-巨球蛋白是蛋白复合物形式的血淋巴的主要成分。我们的结果为黄曲霉血淋巴效应物的进一步功能研究提供了宝贵的资源，可能对tick虫的管理有用。
■通过截肢从完全充血的H.flava蜱中收集血淋巴。通过蓝色天然聚丙烯酰胺凝胶电泳(BN-PAGE)和十二烷基硫酸钠PAGE(SDS-PAGE)检查血淋巴蛋白。通过液相色谱-串联质谱(LC-MS/MS)进一步鉴定从凝胶中提取的蛋白质。
■通过BN-PAGE从蜱血淋巴中分离出两条带(380和520kDa),并通过SDS-PAGE进一步分离成四条带(105、120、130和360kDa)。LC-MS/MS显示在4个条带中存在7个蜱蛋白和13个宿主蛋白。这些蜱蛋白主要属于卵黄蛋白原(Vg)家族和α-巨球蛋白家族成员。计算机结构分析表明，这些Vg家族成员都具有共同的保守结构域，包括N末端脂质结合域（LPD-N），C端vonWillebrandD型结构域(vWD)，和未知函数域(DUF)。此外,通过分析独特的N-末端氨基酸序列和切割位点,确定Vg家族蛋白中的两种属于载体蛋白(CP)。
■这些发现表明，Vg家族蛋白和α-巨球蛋白是蛋白复合物形式的血淋巴的主要成分。我们的结果为黄曲霉血淋巴效应物的进一步功能研究提供了宝贵的资源，可能对tick虫的管理有用。
