光谱学与光谱分析
光譜學與光譜分析
광보학여광보분석
SPECTROSCOPY AND SPECTRAL ANALYSIS
2015年
2期
409-414
,共6页
Ⅰ型胶原溶液%温度%同步荧光光谱%二维相关光谱%聚集行为
Ⅰ型膠原溶液%溫度%同步熒光光譜%二維相關光譜%聚集行為
Ⅰ형효원용액%온도%동보형광광보%이유상관광보%취집행위
Type I collagen%Temperature%Synchronous fluorescence spectroscopy%Two-dimensional correlation spectroscopy%Aggregation behavior
采用恒波长同步荧光法和二维相关分析技术研究了不同浓度Ⅰ型胶原溶液中胶原分子聚集行为随温度升高(10~70℃)的变化规律。选取0.2,0.4,1.6 mg·mL-1的胶原溶液,在初始温度下各浓度溶液中胶原分子分别处于单分子状态、较低程度和较高程度的聚集态。研究表明:波长差为9 nm的同步荧光光谱中,激发波长282和292 nm处荧光峰分别归属于未参与形成氢键的 Tyr(酪氨酸)残基和参与形成氢键的Tyr残基。对升温过程同步荧光数据进行二维相关分析,得两荧光值对温度的响应顺序,进而推测得到:当温度低于30℃时,0.2 mg·mL-1溶液中出现了胶原分子间形成 Tyr 残基参与的氢键的趋势。0.4和1.6 mg·mL-1的溶液中原有聚集体可能发生进一步聚集,形成疏水微区。当逐步接近胶原变性温度(36~38℃)时,推测0.4和1.6 mg·mL-1胶原溶液中的疏水微区和聚集体有被破坏的趋势,而0.2 mg·mL-1胶原溶液保持分子间形成氢键的趋势。超过胶原变性温度时,各浓度溶液中胶原分子三股螺旋结构发生松散。当超过45℃时,胶原分子三股螺旋结构松散的趋势更为明显。
採用恆波長同步熒光法和二維相關分析技術研究瞭不同濃度Ⅰ型膠原溶液中膠原分子聚集行為隨溫度升高(10~70℃)的變化規律。選取0.2,0.4,1.6 mg·mL-1的膠原溶液,在初始溫度下各濃度溶液中膠原分子分彆處于單分子狀態、較低程度和較高程度的聚集態。研究錶明:波長差為9 nm的同步熒光光譜中,激髮波長282和292 nm處熒光峰分彆歸屬于未參與形成氫鍵的 Tyr(酪氨痠)殘基和參與形成氫鍵的Tyr殘基。對升溫過程同步熒光數據進行二維相關分析,得兩熒光值對溫度的響應順序,進而推測得到:噹溫度低于30℃時,0.2 mg·mL-1溶液中齣現瞭膠原分子間形成 Tyr 殘基參與的氫鍵的趨勢。0.4和1.6 mg·mL-1的溶液中原有聚集體可能髮生進一步聚集,形成疏水微區。噹逐步接近膠原變性溫度(36~38℃)時,推測0.4和1.6 mg·mL-1膠原溶液中的疏水微區和聚集體有被破壞的趨勢,而0.2 mg·mL-1膠原溶液保持分子間形成氫鍵的趨勢。超過膠原變性溫度時,各濃度溶液中膠原分子三股螺鏇結構髮生鬆散。噹超過45℃時,膠原分子三股螺鏇結構鬆散的趨勢更為明顯。
채용항파장동보형광법화이유상관분석기술연구료불동농도Ⅰ형효원용액중효원분자취집행위수온도승고(10~70℃)적변화규률。선취0.2,0.4,1.6 mg·mL-1적효원용액,재초시온도하각농도용액중효원분자분별처우단분자상태、교저정도화교고정도적취집태。연구표명:파장차위9 nm적동보형광광보중,격발파장282화292 nm처형광봉분별귀속우미삼여형성경건적 Tyr(락안산)잔기화삼여형성경건적Tyr잔기。대승온과정동보형광수거진행이유상관분석,득량형광치대온도적향응순서,진이추측득도:당온도저우30℃시,0.2 mg·mL-1용액중출현료효원분자간형성 Tyr 잔기삼여적경건적추세。0.4화1.6 mg·mL-1적용액중원유취집체가능발생진일보취집,형성소수미구。당축보접근효원변성온도(36~38℃)시,추측0.4화1.6 mg·mL-1효원용액중적소수미구화취집체유피파배적추세,이0.2 mg·mL-1효원용액보지분자간형성경건적추세。초과효원변성온도시,각농도용액중효원분자삼고라선결구발생송산。당초과45℃시,효원분자삼고라선결구송산적추세경위명현。
The synchronous fluorescence spectroscopy and two dimensional correlation analysis method were applied to study the aggregation behavior of acid-soluble collagen solutions (0. 2,0. 4 and 1. 6 mg·mL-1 )during the heating process of 10~70 ℃. It was found that the fluorescence excited at 292 and 282 nm (Δλ= 9 nm)belongs to the tyrosine (Tyr)residues which partici-pate in forming hydrogen bonds or not,respectively.The two dimensional correlation analysis with the temperature varying showed that with the temperature increased (10~30 ℃)hydrogen bonds among collagen molecular with Tyr residues formed in the 0. 2 mg·mL-1 collagen solution,while the higher aggregations of collagen molecular and hydrophobic micro-domains ap-peared in the 0. 4 and 1. 6 mg·mL-1 collagen solutions.With approaching the denatured temperature of collagen (36~38 ℃), the hydrophobic micro-domain and aggregates seemed to be broken in the 0. 4 and 1. 6 mg·mL-1 collagen solutions,however the hydrogen bonds in the 0. 2 mg·mL-1 were stable.Above the denaturation temperature of collagen,the triple-helix structure of collagen molecular in solution of each concentration tended to be loose.In the heating process of 45~70 ℃,this trend was more obvious.