Detection of constitutive heterodimerization of the integrin Mac-1 subunits by fluorescence resonance energy transfer in living cells

Journal Article · Fri Aug 04 00:00:00 EDT 2006 · Biochemical and Biophysical Research Communications

OSTI ID:20854395

Guo, Fu ^[1]; Huayan, Yang ^[2]; Chen, Wang ^[1]; Feng, Zhang ^[2]; Zhendong, You ^[2]; Guiying, Wang ^[1]; Cheng, He ^[2]; Yizhang, Chen ^[2]; Zhihan, Xu ^[1]

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1. State Key Laboratory of High Field Laser Physics, Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, Shanghai 201800 (China)
2. Institute of Neuroscience, Department of Neurobiology, Second Military Medical University, Shanghai 200433 (China)

Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to {beta}{sub 2} subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively.

• OSTI ID: 20854395
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 3 Vol. 346; ISSN 0006-291X; ISSN BBRCA9
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ANTIGENS
ENERGY TRANSFER
FLUORESCENCE
FLUORESCENCE SPECTROSCOPY
HAMSTERS
IN VITRO
IN VIVO
KIDNEYS
LEUKOCYTES
MACROPHAGES
MAXIMUM ACCEPTABLE CONTAMINATION
MICROSCOPY
OVARIES
RECEPTORS