tailieunhanh - Báo cáo khoa học: Purification and characterization of zebrafish hatching enzyme – an evolutionary aspect of the mechanism of egg envelope digestion

There are two hatching enzyme homologues in the zebrafish genome: zebrafish hatching enzymeZHE1andZHE2. Northern blot and RT-PCR analysis revealed that ZHE1 was mainly expressed in pre-hatching embryos, whereasZHE2was rarely expressed. This was consistent with the results obtained in an experiment conducted at the protein level, which demonstrated that one kind of hatching enzyme, ZHE1, was able to be purified from the hatching liquid. | Purification and characterization of zebrafish hatching enzyme - an evolutionary aspect of the mechanism of egg envelope digestion Kaori Sano1 Keiji Inohaya2 Mari Kawaguchi3 Norio Yoshizaki4 Ichiro luchi5 and Shigeki Yasumasu5 1 Graduate Program of BiologicalScience Graduate Schoolof Science and Technology Sophia University Tokyo Japan 2 Department of BiologicalInformation Tokyo Institute of Technology Yokohama Japan 3 Ocean Reseach Institute The University of Tokyo Japan 4 Department of BiologicalDiversity Faculty of Agriculture Gifu University Japan 5 Department of Materials and Life Sciences Faculty of Science and Technology Sophia University Tokyo Japan Keywords astacin family egg envelope hatching enzyme molecular evolution Correspondence S. Yasumasu Department of Materials and Life Sciences Faculty of Science and Technology Sophia University 7-1 Kioi-cho Chiyoda-ku Tokyo 102-8554 Japan Fax Tel 81 3 3238 3393 E-mail s-yasuma@ Received 17 June 2008 revised 22 September 2008 accepted 2 October 2008 doi There are two hatching enzyme homologues in the zebrafish genome zebrafish hatching enzyme ZHE1 and ZHE2. Northern blot and RT-PCR analysis revealed that ZHE1 was mainly expressed in pre-hatching embryos whereas ZHE2 was rarely expressed. This was consistent with the results obtained in an experiment conducted at the protein level which demonstrated that one kind of hatching enzyme ZHE1 was able to be purified from the hatching liquid. Therefore the hatching of zebrafish embryo is performed by a single enzyme different from the finding that the medaka hatching enzyme is an enzyme system composed of two enzymes medaka high choriolytic enzyme MHCE and medaka low chorio-lytic enzyme MLCE which cooperatively digest the egg envelope. The six ZHE1-cleaving sites were located in the N-terminal regions of egg envelope subunit proteins ZP2 and ZP3 but not in the internal regions such as the ZP domains. The digestion .

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