Abstract:
:A calcium activated photoprotein, termed mnemiopsin, which emits bioluminescence upon the addition of calcium ion, has been isolated from the Ctenophore, Memiopsis leidyi, and purified by hollow fiber techniques. The system is similar to aequorin, from the jellyfish Aequorea, except that mnemiopsin can be light-inactivated. Separation of mnemiopsin from the dilute and large volume animal homogenate proved difficult with conventional biochemical techniques. A continuous flow process utilizing large surface area hollow fibers for filtration, concentration, and dialysis was developed which may also be applicable to the purification of other proteins. The resulting mnemiopsin concentrate, after further purification, was judged to be about 90% pure by its gel electrophoretic profile. Estimates by molecular sieve chromatography and SDS gel electrophoresis gave a molecular weight of about 23,000 daltons. A calcium specificity for triggering light emission was studied by comparison of triggering with a variety of cations and anions and by investigating the effects of calcium ionophores and antagonists. The activity of mnemiospin was characterized with respect to pH, temperature and ionic strength. The stability of mnemiopsin activity after exposure to proteases, denaturants, protein group specific reagents, detergents, elevated temperatures and light was determined. Some years ago our laboratory reported that the bioluminescence reaction in the ctenophores which had long eluded definition involved a calcium activated photoprotein similar in many respects to that found in other coelenterates, notably Aequorea. We found, moreover, that the systems differed in that the bioluminescent activity of the isolated protein was lost following exposure to light. The purification and characterization of this biochemical system was undertaken both in our laboratory and by Ward and Seliger. These latter reports provide a detailed and firm foundation for the understanding of the components and mechanisms involved. While many of our results are in agreement with theirs, our approaches, inquiries, and results differed in several significant ways, the description of which forms the basis for this report. In particular, we took a different approach in the purification of the Mnemiopsis photoprotein which in itself is rather a formidable task. The technique was successful and may point the way to other applications where large volume dilute solutions prove cumbersome. Secondly, our study of the effects of salts, proteases, detergents, and other agents indicate that the protein, though sensitive to calcium and visible light inactivation, is relatively resistant to some agents which commonly inactivate proteins.
journal_name
Mol Cell Biochemjournal_title
Molecular and cellular biochemistryauthors
Girsch SJ,Hastings JWdoi
10.1007/BF00232600subject
Has Abstractpub_date
1978-04-11 00:00:00pages
113-24issue
2eissn
0300-8177issn
1573-4919journal_volume
19pub_type
杂志文章abstract::Administration of nifedipine to mice over a period of six months caused a significant (p < 0.05) decrease in neutrophilic functions viz superoxide generation, coupled to NADPH oxidase activity as well as NADPH production by HMP shunt. Properties like chemotaxis and phagocytosis showed a similar decrease. From this stu...
journal_title:Molecular and cellular biochemistry
pub_type: 杂志文章
doi:10.1007/BF00925987
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journal_title:Molecular and cellular biochemistry
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journal_title:Molecular and cellular biochemistry
pub_type: 杂志文章
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journal_title:Molecular and cellular biochemistry
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journal_title:Molecular and cellular biochemistry
pub_type: 杂志文章
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pub_type: 杂志文章,评审
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doi:10.1007/s11010-009-0300-5
更新日期:2010-04-01 00:00:00
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journal_title:Molecular and cellular biochemistry
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doi:
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journal_title:Molecular and cellular biochemistry
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更新日期:2000-04-01 00:00:00
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doi:10.1007/s11010-017-3070-5
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journal_title:Molecular and cellular biochemistry
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doi:10.1007/BF01741714
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journal_title:Molecular and cellular biochemistry
pub_type: 杂志文章
doi:10.1007/BF01076564
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journal_title:Molecular and cellular biochemistry
pub_type: 杂志文章
doi:10.1007/BF02357031
更新日期:1981-05-26 00:00:00
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journal_title:Molecular and cellular biochemistry
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journal_title:Molecular and cellular biochemistry
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pub_type: 杂志文章
doi:10.1023/a:1006851426257
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journal_title:Molecular and cellular biochemistry
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pub_type: 杂志文章
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更新日期:2008-01-01 00:00:00
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journal_title:Molecular and cellular biochemistry
pub_type: 杂志文章
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更新日期:2014-08-01 00:00:00
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pub_type: 杂志文章
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