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电子传递黄素蛋白小亚基与通用酰基辅酶A脱氢酶的优先交联。

Preferential cross-linking of the small subunit of the electron-transfer flavoprotein to general acyl-CoA dehydrogenase.

作者信息

Steenkamp D J

出版信息

Biochem J. 1987 Apr 15;243(2):519-24. doi: 10.1042/bj2430519.

DOI:10.1042/bj2430519
PMID:3115254
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1147885/
Abstract

The interaction between pig liver mitochondrial electron-transfer flavoprotein (ETF) and general acyl-CoA dehydrogenase (GAD) was investigated by means of the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. Neither ETF or GAD contained reactive thiol groups. The substitution of 9.4 lysine residues/FAD group in GAD with pyridyl disulphide structures did not affect the catalytic activity of the enzyme. Thiol groups were introduced into ETF by thiolation with methyl 4-mercaptobutyrimidate. ETF containing 10.5 reactive thiol groups/FAD group showed undiminished electron-acceptor activity with respect to GAD. The reaction of thiolated ETF and GAD containing pyridyl disulphide structures resulted in a decreased staining intensity of the small subunit of ETF on SDS/polyacrylamide-gel electrophoresis. Preferential cross-linking of the smaller subunit of ETF to GAD did not take place when ETF was first treated with SDS, but was unaffected by reduction of GAD by octanoyl-CoA.

摘要

通过异双功能试剂N-琥珀酰亚胺基3-(2-吡啶二硫代)丙酸酯研究了猪肝线粒体电子传递黄素蛋白(ETF)与一般酰基辅酶A脱氢酶(GAD)之间的相互作用。ETF和GAD均不含有反应性巯基。用吡啶二硫结构取代GAD中9.4个赖氨酸残基/FAD基团不会影响该酶的催化活性。通过用4-巯基丁酸甲酯硫醇化将巯基引入ETF。含有10.5个反应性巯基/FAD基团的ETF对GAD表现出未减弱的电子受体活性。含有吡啶二硫结构的硫醇化ETF与GAD的反应导致SDS/聚丙烯酰胺凝胶电泳上ETF小亚基的染色强度降低。当ETF先用SDS处理时，ETF较小亚基与GAD的优先交联不会发生，但不受辛酰辅酶A对GAD还原的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/9faef6c7b919/biochemj00257-0211-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/b6b21ad0f482/biochemj00257-0210-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/21f997075949/biochemj00257-0210-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/9faef6c7b919/biochemj00257-0211-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/b6b21ad0f482/biochemj00257-0210-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/21f997075949/biochemj00257-0210-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70c4/1147885/9faef6c7b919/biochemj00257-0211-a.jpg

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Cross-linking of the electron-transfer flavoprotein to electron-transfer flavoprotein-ubiquinone oxidoreductase with heterobifunctional reagents.利用异双功能试剂将电子传递黄素蛋白与电子传递黄素蛋白-泛醌氧化还原酶交联。
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