Abstract:
:Galactocerebrosidase was purified about 22,600-fold using several hydrophobic column and gel filtration steps with a 4.8% recovery, from human lymphocytes. Its specific activity was 1.54 x 10(5) nmol/h/mg with tritium-labeled galactocerebroside as the substrate in the taurocholate system. The optimal pH for galactocerebroside was 4.2 in the taurocholate system and 4.6 in the cholate system. The Km values for galactocerebroside were 5 microM in the taurocholate system and 25 microM in the cholate system. The molecular weight of the purified enzyme was estimated to be 90 kDa by sodium dodecylsulfate polyacrylamide gel electrophoresis and gel filtration. However, 70, 50, 40, and 30 kDa bands were also recognized on SDS-PAGE. The N-terminal amino acid sequences of the 70 kDa molecule and the three 50 kDa molecules were the same as that of the 90 kDa molecule. The N-terminal amino acid sequences of the 40 and 30 kDa molecules were unique. A monoclonal antibody raised against the purified enzyme effectively immunoprecipitated galactocerebrosidase activity, and an affinity column prepared with this monoclonal antibody bound the 90 and 50 kDa proteins. These results suggest that this enzyme is probably processed from the 90 kDa protein.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Sakai N,Inui K,Midorikawa M,Okuno Y,Ueda S,Iwamatsu A,Okada Sdoi
10.1093/oxfordjournals.jbchem.a124569subject
Has Abstractpub_date
1994-09-01 00:00:00pages
615-20issue
3eissn
0021-924Xissn
1756-2651journal_volume
116pub_type
杂志文章abstract::The gene encoding a MutM from Desulfovibrio vulgaris (Miyazaki F) was cloned and expressed in Escherichia coli. A 5.9-kb DNA fragment, isolated from D. vulgaris (Miyazaki F) by XhoI and PvuII, contained a MutM gene and other open reading frames. The nucleotide sequence of the MutM gene indicated that the protein was c...
journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:2009-04-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134586
更新日期:1984-01-01 00:00:00
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journal_title:Journal of biochemistry
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更新日期:1998-10-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1998-07-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a132589
更新日期:1979-09-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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journal_title:Journal of biochemistry
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更新日期:1980-02-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
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更新日期:1985-06-01 00:00:00
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更新日期:2009-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:1988-02-01 00:00:00
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journal_title:Journal of biochemistry
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更新日期:2002-12-01 00:00:00
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