Structural Characterization of Fish Egg Vitelline Envelope Proteins by Mass Spectrometry
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- 作者：Costel C. Darie ; Martin L. Biniossek ; Luca Jovine ; Eveline S. Litscher ; and Paul M. Wassarman
- 刊名：Biochemistry
- 出版年：2004
- 出版时间：June 15, 2004
- 年：2004
- 卷：43
- 期：23
- 页码：7459 - 7478
- 全文大小：964K
- 年卷期：v.43,no.23(June 15, 2004)
- ISSN：1520-4995
文摘
The extracellular coat, or vitelline envelope (VE), of rainbow trout (Oncorhynchus mykiss)eggs consists of three proteins, called VE
lpha.gif" BORDER=0> (Mr ~52 kDa), VE
le"> (Mr ~48 kDa), and VE
(Mr ~44 kDa).Each of these proteins is related to mammalian egg zona pellucida (ZP) glycoproteins ZP1-3 and possessesan N-terminal signal sequence, a ZP domain, and a protease cleavage site near the C-terminus. VElpha.gif" BORDER=0> andVEle"> also have a trefoil domain. All three proteins possess a relatively large number of cysteine residues(VElpha.gif" BORDER=0>, 18; VEle">, 18; VE, 12), of which 8 are present in the ZP domain and 6 are present in the trefoildomain of VElpha.gif" BORDER=0> and VEle">. Here, several types of mass spectrometry were employed, together with gelelectrophoresis of chemical and enzymatic digests, to identify intramolecular disulfide linkages, as wellas the N- and C-terminal amino acids of VElpha.gif" BORDER=0>, VEle">, and VE. Additionally, these methods were used tocharacterize two high molecular weight proteins (HMWPs; Mr > 110 kDa) of rainbow trout VEs that areheterodimers of individual VE proteins. These analyses have permitted assignment of disulfide linkagesand identification of N- and C-terminal amino acids for the VE proteins and determination of the proteincomposition of two forms of HMWPs. These experiments provide important structural information aboutfish egg VE proteins and filaments and about structural relationships between extracellular coat proteinsof mammalian and nonmammalian eggs.
lpha.gif" BORDER=0> (Mr ~52 kDa), VEle"> (Mr ~48 kDa), and VE (Mr ~44 kDa).Each of these proteins is related to mammalian egg zona pellucida (ZP) glycoproteins ZP1-3 and possessesan N-terminal signal sequence, a ZP domain, and a protease cleavage site near the C-terminus. VElpha.gif" BORDER=0> andVEle"> also have a trefoil domain. All three proteins possess a relatively large number of cysteine residues(VElpha.gif" BORDER=0>, 18; VEle">, 18; VE, 12), of which 8 are present in the ZP domain and 6 are present in the trefoildomain of VElpha.gif" BORDER=0> and VEle">. Here, several types of mass spectrometry were employed, together with gelelectrophoresis of chemical and enzymatic digests, to identify intramolecular disulfide linkages, as wellas the N- and C-terminal amino acids of VElpha.gif" BORDER=0>, VEle">, and VE. Additionally, these methods were used tocharacterize two high molecular weight proteins (HMWPs; Mr > 110 kDa) of rainbow trout VEs that areheterodimers of individual VE proteins. These analyses have permitted assignment of disulfide linkagesand identification of N- and C-terminal amino acids for the VE proteins and determination of the proteincomposition of two forms of HMWPs. These experiments provide important structural information aboutfish egg VE proteins and filaments and about structural relationships between extracellular coat proteinsof mammalian and nonmammalian eggs.