Abstract：
:The ubiquitin-proteasome system is essential for intracellular protein degradation, but there are few studies of this system in the extracellular milieu. Recently, we reported that a 70-kDa sperm receptor, HrVC70, on the vitelline coat is ubiquitinated and then degraded by the sperm proteasome during fertilization of the ascidian, Halocynthia roretzi. Here, we investigated the mechanism of extracellular ubiquitination. The HrVC70-ubiquitinating enzyme activity was found to be released from the activated sperm during the fertilization process. This enzyme was purified from an activated sperm exudate, by chromatography on DEAE-cellulose and ubiquitin-agarose columns, and by glycerol density gradient centrifugation. The molecular mass of the enzyme was estimated to be 700 kDa. The purified enzyme requires CaCl2 and MgATP for activity, and is active in seawater. The purified enzyme preparation, but not the crude enzyme preparation, showed narrow substrate specificity to HrVC70. Moreover, ATP and ubiquitin are released from the activated sperm to the surrounding seawater during fertilization. These results indicate that ascidian sperm release a novel extracellular ubiquitinating enzyme system together with ATP and ubiquitin during penetration of the vitelline coat of the egg, which catalyzes the ubiquitination of the HrVC70, an essential component of ascidian fertilization.
journal_name
Dev Bioljournal_title
Developmental biologyauthors
Sakai N,Sawada H,Yokosawa Hdoi
10.1016/j.ydbio.2003.07.016subject
Has Abstractpub_date
2003-12-01 00:00:00pages
299-307issue
1eissn
0012-1606issn
1095-564Xpii
S0012160603004500journal_volume
264pub_type
杂志文章abstract：:Although mitogen-activated protein kinase (MAPK) is a well-known cell cycle regulator, emerging studies have also implicated its activity in the regulation of intracellular calcium concentration ([Ca2+](i)) and secretion. Those studies raise the hypothesis that MAPK activity during oocyte maturation and early fertiliz...
journal_title：Developmental biology
pub_type： 杂志文章
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journal_title：Developmental biology
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journal_title：Developmental biology
pub_type： 杂志文章,评审
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journal_title：Developmental biology
pub_type： 杂志文章
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pub_type： 杂志文章
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pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2008-04-15 00:00:00
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journal_title：Developmental biology
pub_type： 杂志文章
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pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2006-02-15 00:00:00
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journal_title：Developmental biology
pub_type： 杂志文章
doi：10.1016/j.ydbio.2020.09.008
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journal_title：Developmental biology
pub_type： 杂志文章
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更新日期：1989-12-01 00:00:00
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pub_type： 杂志文章
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更新日期：2017-09-01 00:00:00
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journal_title：Developmental biology
pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2003-06-15 00:00:00
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journal_title：Developmental biology
pub_type： 杂志文章
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更新日期：2000-11-01 00:00:00
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pub_type： 杂志文章
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更新日期：2016-07-15 00:00:00
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journal_title：Developmental biology
pub_type： 杂志文章
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pub_type： 杂志文章
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