Dynamic atomic force microscopic observation of the three-dimensional conformation of protein A-gold single molecule

Abstract

Abstract: Objective To determine the three-dimensional(3D) conformation of staphylococcal protein A-gold(SPA-G) single molecule using atomic force microscope(AFM) so as to evaluate the feasibility of using this molecule for in situ labeling of the neuronal membrane protein. Methods AFM was used to acquire the images of SPA-G binding to the surface of mica under physiological condition for determining the 3D conformation of the molecule. Results SPA-G single molecule was shown to contain a characteristic structure with a chain in the shape of mirror image of the letter C, which had the dimension of 48.80 nm ×42.13 nm×20.53 nm. Conclusion AFM provide a new means for morphological investigation of the biomacromolecule at the nanometer scale in physiological conditions, and SPA-G can be utilized for in situ labeling of the neuronal membrane receptors.

CLC Number:

Q446.8

YU Yi-gang, XU Ru-xiang, CAI Ying-qian, JIANG Xiao-dan, KE Yi-quan. Dynamic atomic force microscopic observation of the three-dimensional conformation of protein A-gold single molecule[J]. Journal of Southern Medical University, 2005, 25(02): 139-142.

References

[1] Yamakura T, Shimoji K. Subunit-and site-specific pharmacology of the NMDA receptor channel [J]. ProgNeurobiol, 1999, 59(3):279-98.
[2] 赵雨杰.医学生物信息学[M]. 北京: 人民军医出版社, 2002.4-5.
[3] Sjoholm I. Protein A from Staphylococcus aureus. Spectropolarimetric and spectrophotometric studies [J]. Eur J Biochem, 1975, 51(1): 55-61.
[4] MacKintosh FR, Bennett K, Shiff S, et al. Treatment of advanced malignancy with plasma perfused over staphylococcal protein A [J].West J Med, 1983, 139(1): 36-40.
[5] Ohnishi S, Murata M, Hato M, et al. Correlation between surfacemorphology and surface force of protein A adsorbed on mica [J].Biophys J, 1998, 74(1): 455-65.
[6] 郁毅刚,徐如祥.纳米技术在神经外科中应用展望[J].国外医学·神经病学与神经外科分册(Foreign Med Sci·Neurosurg NeurolSect),2001,28(5):345-7.
[7] Le Grimellec C, Lesniewska E, Cachia C, et al. Imaging of the membrane surface of MDCK cells by atomic force microscopy [J]. Biophys J, 1994, 67(1): 36-41.
[8] Utsuno K, Tsuboi M, Katsumata S, et al. Viewing of complex molecules of ethidium bromide and plasmid DNA in solution by atomic microscopy[J]. Chem Pharm Bull, 2001, 49(4): 413-7.
[9] Shlyakhtenko LS, Gall AA, Weimer JJ, et al. Atomic force microscopy imaging of DNA covalently immobilized on a functionalised mica substrate[J]. Biophys J, 1999, 77(1): 568-76.
[10] Binnig G, Quate CF, Gerber C. Atomic force microscope [J]. Phys Rew Lett, 1986, 56(9): 930-3.
[11] Hermann R, Walther P, Mueller M. Immunogold labeling in scan ning electron microscopy [J]. Histochem Cell Biol, 1996, 106(3):356-9.
[12] Bustamante C, Vesonka J, Tang CL, et al. Circular DNA molecules imaged in air by scanning force microscopy[J]. Biochem, 1992, 31(1): 22-6.
[13] Xu S, Arusdorf MF. Calibration of the scanning(atomic) force microscope with gold particles [J]. J Microsc, 1994, 173(Pt 3): 199-210.
[14] Sjodahl J. Structural studies on the four repetitive Fc-binding regions in protein A from Staphylococcus aureus [J]. Eur J Biochem, 1977,78(2): 471-9.
[15] Ouerghi O, Touhami A, Othnane A, et al. Investigating antibodyantigen binding with atomic force microscopy [J]. Biomol Eng,2002, 19(2-6): 183-8.