海洋科学2024，Vol.48Issue(2)：24-31,8.DOI:10.11759/hykx20220924001
采用SDS研究泥蚶血红蛋白(Tg-HbⅡ)具有过氧化物酶及抗菌活性的结构基础
Investigation of the structural basis for peroxidase and anti-bacterial activity of hemoglobin from blood clam Tegillarca granosa(Tg-HbⅡ)using sodium dodecyl sulfate
摘要
Abstract
Hemoglobin Ⅱ from Tegillarca granosa(Tg-HbⅡ)possesses oxygen-carrying capabilities as well as peroxidase and antibacterial activities.In this study,we examined the effects of sodium dodecyl sulfate(SDS)on the structure,peroxidase activity,and antibacterial activity of Tg-HbⅡ using spectrophotometric,spectroscopic,and molecular docking techniques.Moreover,we explored the structural basis of the peroxidase activity of Tg-HbⅡand its antibacterial activities.The results revealed that the hydrophobic alkyl long chains of SDS could integrate into the heme pocket of Tg-HbⅡ,forming hydrogen bonds with the proximal His104.This interaction resulted in the breaking of the coordination bond between heme iron and His104,leading to a decrease and shift in the Soret band absorption peak of Tg-HbⅡ.Additionally,SDS could form hydrophobic interactions with the amino acids in the heme pocket,inducing alterations in the original structure and exposing some hydrophobic amino acids.This re-sulted in an enhanced exogenous fluorescence intensity and a redshift of the maximum emission wavelength.SDS demonstrated an inhibitory effect on the peroxidase activity of Tg-HbⅡ.At 2 mmol·L-1 of SDS concentration,the enzymatic activity of Tg-HbⅡ was only 20％of that of the original,and the antibacterial activity against Bacillus subtilis was lost in agar diffusion experiments.These results indicated that SDS inhibited the peroxidase activity of Tg-HbⅡ by disrupting the internal structure of the heme pocket,consequently leading to the loss of its antibacterial activity.The heme hydrophobic pocket emerged as a key structure for the peroxidase and antibacterial activities of Tg-HbⅡ.This study lays the foundation for future studies on the antibacterial mechanism of Tg-HbⅡ.关键词
泥蚶血红蛋白/十二烷基硫酸钠/过氧化物酶活性/抗菌活性/血红素口袋Key words
Tegillarca granosa hemoglobin/sodium dodecyl sulfate/peroxidase activity/antibacterial activity/heme pocket分类
生物科学引用本文复制引用
濮丽丽,王诗怡,林志华,王素芳,包永波..采用SDS研究泥蚶血红蛋白(Tg-HbⅡ)具有过氧化物酶及抗菌活性的结构基础[J].海洋科学,2024,48(2):24-31,8.基金项目
浙江省重点自然科学基金项目(LZ20C190001) （LZ20C190001）
浙江省科技重大专项(2021C02069-7) （2021C02069-7）
宁波市公益研究重点项目(2021S014) （2021S014）
浙江省生物工程重中之重学科项目(CX2021001)和浙江省高校基本科研业务费项目[Key Natural Science Foundation of Zhejiang Province,No.LZ20C190001 （CX2021001）
Major Science and Technology Projects of Zhejiang Prov-ince,No.2021C02069-7 （）
Ningbo Public Benefit Research Key Project,No.2021S014 （）
Zhejiang Provincial Top Key Discipline of Biological Engi-neering,No.CX2021001,and Basic Scientific Research Operating Ex-pense Project of Zhejiang Provincial Universities] （）
