Rajamoorthi K, Brown M F
Department of Chemistry, University of Arizona, Tucson 85721.
Biochemistry. 1991 Apr 30;30(17):4204-12. doi: 10.1021/bi00231a015.
The configurational properties and dynamics of the arachidonic acyl chains of phospholipid bilayers have been investigated for the first time by solid-state 2H NMR techniques, with the goal of achieving a better understanding of the biological roles of polyunsaturated phospholipids. Vinyl perdeuterated arachidonic acid (20:4 delta 5,8,11,14-d8) was prepared from eicosatetraynoic acid (ETYA) and was esterified with 1-palmitoyl-sn-glycero-3-phosphocholine to yield 1-palmitoyl-2-vinylperdeuterioarachidonoyl-sn-glycero-3-phosphocho line [(16:0)(20:4-d8)PC]. 31P NMR spectra of aqueous dispersions of (16:0)(20:4-d8)PC as well as 1-perdeuteriopalmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine [(per-2H-16:0)(20:4)PC] were characteristic of the lamellar liquid-crystalline state. The dispersions had similar 31P chemical shift anisotropies, with little apparent motional averaging of the lineshapes due to macroscopic reorientation of liposomes or lateral diffusion of phospholipids about their curved surfaces. Comparison to other phosphatidylcholines indicated that both samples comprised the fully hydrated L alpha phase plus excess water. However, the dispersion of (16:0)(20:4-d8)PC yielded relatively narrow powder-type 2H NMR spectra, compared to (per-2H-16:0)(20:4)PC in the liquid-crystalline state. The differences in the 2H NMR powder patterns thus reflect differences in the configurational properties of the polyunsaturated sn-2 arachidonic acyl chain compared to the saturated sn-1 palmitic chain. When the powder-type 2H NMR spectra of the (16:0)(20:4-d8)PC bilayer were dePaked (theta = 0 degrees), they showed three kinds of deuterons upon integration: one with a large splitting (approximately 25-35 kHz), two with intermediate splittings (approximately 10-15 kHz), and the remainder with smaller splittings (approximately 0.3-5 kHz).(ABSTRACT TRUNCATED AT 250 WORDS)
首次采用固态2H NMR技术研究了磷脂双层中花生四烯酸酰基链的构型性质和动力学,目的是更好地理解多不饱和磷脂的生物学作用。乙烯基全氘代花生四烯酸(20:4δ5,8,11,14-d8)由二十碳四炔酸(ETYA)制备,并与1-棕榈酰-sn-甘油-3-磷酸胆碱酯化,得到1-棕榈酰-2-乙烯基全氘代花生四烯酰-sn-甘油-3-磷酸胆碱[(16:0)(20:4-d8)PC]。(16:0)(20:4-d8)PC以及1-全氘代棕榈酰-2-花生四烯酰-sn-甘油-3-磷酸胆碱[(全-2H-16:0)(20:4)PC]水分散体的31P NMR谱具有层状液晶态的特征。这些分散体具有相似的31P化学位移各向异性,由于脂质体的宏观重排或磷脂在其弯曲表面的横向扩散,线形状几乎没有明显的运动平均化。与其他磷脂酰胆碱相比表明,两个样品均包含完全水合的Lα相和过量的水。然而,与液晶态的(全-2H-16:0)(20:4)PC相比,(16:0)(20:4-d8)PC的分散体产生相对较窄的粉末型2H NMR谱。因此,2H NMR粉末图谱的差异反映了与饱和sn-1棕榈酸链相比,多不饱和sn-2花生四烯酸酰基链构型性质的差异。当(16:0)(20:4-d8)PC双层的粉末型2H NMR谱解包(θ = 0度)时,积分后显示出三种氘核:一种具有大的分裂(约25-35 kHz),两种具有中等分裂(约10-15 kHz),其余具有较小的分裂(约0.3-5 kHz)。(摘要截短于250字)