JP2895097B2 - Method for preparing protein two-dimensional crystal film - Google Patents
Method for preparing protein two-dimensional crystal filmInfo
- Publication number
- JP2895097B2 JP2895097B2 JP18804989A JP18804989A JP2895097B2 JP 2895097 B2 JP2895097 B2 JP 2895097B2 JP 18804989 A JP18804989 A JP 18804989A JP 18804989 A JP18804989 A JP 18804989A JP 2895097 B2 JP2895097 B2 JP 2895097B2
- Authority
- JP
- Japan
- Prior art keywords
- film
- liquid
- dimensional crystal
- liquid film
- crystal film
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
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- Application Of Or Painting With Fluid Materials (AREA)
- Manufacture Of Macromolecular Shaped Articles (AREA)
- Peptides Or Proteins (AREA)
Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は蛋白２次元結晶膜を液液界面において作成す
る方法に関するものである。The present invention relates to a method for forming a protein two-dimensional crystal film at a liquid-liquid interface.
（従来の技術） 従来、蛋白２次元結晶膜を作成するにあたっては、２
次元結晶膜作成トラフ内の水面に界面活性剤を使用して
アンカー膜を形成し、このアンカー膜に蛋白分子を吸着
させる。一般にこのアンカー膜を作成する界面活性剤と
してステアリルアミンを使用し、均一膜厚となるよう
に、一定の表面圧になるように調整しているが、横方向
の流動性に欠けるため、均一性に欠け、作成が困難であ
るとともに、２次元結晶膜の結晶性が不十分である。(Prior Art) Conventionally, when producing a protein two-dimensional crystal film, two
An anchor film is formed on a water surface in a trough for forming a two-dimensional crystal film using a surfactant, and protein molecules are adsorbed to the anchor film. Generally, stearylamine is used as a surfactant to form the anchor film, and the surface pressure is adjusted so as to obtain a uniform film thickness. And the two-dimensional crystal film has insufficient crystallinity.
（発明の解決しようとする課題） しかるに、アンカー膜を形成する界面活性剤に横方向
の流動性を付与すべく、コレストロールを添加配合する
ことが提案されるものの、アンカー膜を一定の表面圧に
調整することを必要とし、アンカー膜調製の困難性が残
存するという問題点がある。そこで、本発明は均一な２
次元結晶膜の作成方法を提供することを目的とする。(Problems to be Solved by the Invention) However, although it has been proposed to add and blend cholesterol in order to impart lateral fluidity to the surfactant forming the anchor film, the anchor film is required to have a constant surface pressure. There is a problem that adjustment is required, and the difficulty of preparation of the anchor membrane remains. Therefore, the present invention provides a uniform 2
It is an object to provide a method for forming a two-dimensional crystal film.
（課題を解決するための手段） 本発明は従来のLB膜の作成の問題点が界面活性剤を使
用した気液界面における吸着現象を利用することに起因
するものであり、蛋白分子吸着膜としてデヒドロアビエ
チルアミンおよびその誘導体に代表される第１級アミノ
基を有する横方向流動性に優れる均一な担持膜を形成す
れば、液液界面における吸着によりLB膜を形成すること
ができることに着目してなされたもので、 蛋白質水溶液を撥水性凹部又はトラフ内の膜形成室内
に入れ、該蛋白質水溶液面に、蛋白分子に対し吸着性を
示す第１級アミノ基を有する水に不溶な液体の層からな
る液膜を形成し、該蛋白質水溶液面と該液膜との液液界
面において、該液膜を担持液膜として、該液膜に蛋白分
子を吸着・担持させる工程を含む蛋白２次元結晶膜作成
方法にある。(Means for Solving the Problems) In the present invention, the problem of the conventional preparation of the LB film is due to the use of the adsorption phenomenon at the gas-liquid interface using a surfactant. Focusing on the fact that if a uniform support film having primary amino groups represented by dehydroabiethylamine and its derivatives and having excellent lateral fluidity is formed, an LB film can be formed by adsorption at the liquid-liquid interface. An aqueous protein solution is placed in a water-repellent recess or a film formation chamber in a trough, and a water-insoluble liquid layer having a primary amino group having an adsorptivity to protein molecules is formed on the surface of the aqueous protein solution. A two-dimensional crystal film comprising the steps of: forming a liquid film, and adsorbing and supporting protein molecules on the liquid film at the liquid-liquid interface between the aqueous protein solution surface and the liquid film, using the liquid film as a supporting liquid film. How to make A.
本発明によれば、蛋白質水溶液の表面に形成される担
持膜は横方向流動性に優れる液膜として形成されるの
で、界面活性剤によるアンカー膜と異なり、表面圧を調
整せずとも自動的に均一性に優れたものとなる。According to the present invention, since the carrier film formed on the surface of the aqueous protein solution is formed as a liquid film having excellent lateral fluidity, unlike an anchor film using a surfactant, it is automatically formed without adjusting the surface pressure. It becomes excellent in uniformity.
本発明において使用される担持液膜形成物質は次式で
示されるデヒドロアビエチルアミンまたはその誘導体を
使用するのが好ましい。As the material for forming a supported liquid film used in the present invention, it is preferable to use dehydroabiethylamine or a derivative thereof represented by the following formula.
式中、ｎは１〜４、Ｒは水素または第１級アミノアル
キルを示す。 In the formula, n represents 1-4, and R represents hydrogen or primary aminoalkyl.
かかるデヒドロアビエチルアミンおよびその誘導体は
ファナスチレン環を基本骨格とし、アミノアルキル置換
されていることもあるアミノ基を有することを特徴と
し、種々の点で液液界面における単分子膜形成用アンカ
ー液膜として適当な物性を示す。Such dehydroabiethylamine and its derivatives have a basic skeleton of a fanastyrene ring and have an amino group which may be aminoalkyl-substituted, and in various respects an anchor liquid film for forming a monomolecular film at a liquid-liquid interface Shows suitable physical properties.
本発明において、蛋白質水溶液としては形成する担持
液膜によって異なってよいが、上記液膜物質が不溶であ
る水が通常用いられてよい。In the present invention, the aqueous protein solution may vary depending on the supported liquid film to be formed, but water in which the liquid film substance is insoluble may be usually used.
以下、本発明を添付図面に示す具体例に基づき、詳細
に説明することにする。Hereinafter, the present invention will be described in detail based on specific examples shown in the accompanying drawings.
（実施例） 第１図は本発明方法を適用する単分子膜作成装置の概
要を示す断面説明図で、テフロン樹脂から製造された板
１上面に直径4mm、深さ１〜2mmの平坦な穴からなる収納
部11を複数個形成している。(Example) FIG. 1 is a sectional explanatory view showing an outline of a monomolecular film forming apparatus to which the method of the present invention is applied. A flat hole having a diameter of 4 mm and a depth of 1 to 2 mm is formed on the upper surface of a plate 1 made of Teflon resin. Are formed.
上記単分子膜作成装置の収容部11にはフェリチン0.1m
g/ml（クエン酸緩衝液）濃度の10mmモル液（蛋白質水溶
液）Ｌを導入し、その表面にデヒドロアビエチルアミン
0.1mg/ml（ヘキサン）濃度液を1000Å以上の表面液膜Ｓ
を形成するに必要な量を導入する（第２図参照）。Ferritin 0.1 m in the storage unit 11 of the above monomolecular film forming apparatus
g / ml (citrate buffer) of 10 mm molar solution (protein aqueous solution) L was introduced, and dehydroabiethylamine was added to the surface.
0.1mg / ml (hexane) concentration liquid over 1000 を surface liquid film S
(See FIG. 2).
平衡化させた後、水平付着法にてカーボン膜を貼った
EMメッシュに蛋白２次元結晶膜をサンプリングした。After equilibration, a carbon film was applied by the horizontal attachment method.
The protein two-dimensional crystal film was sampled on the EM mesh.
上記方法で作成した２次元結晶膜は高い結晶性を有し
ていた。The two-dimensional crystal film formed by the above method had high crystallinity.
（発明の効果） 以上の説明で明らかなように、本発明によれば、水面
膜に代表されるアンカー膜を界面活性剤によらず、横方
向流動性に優れる液膜にて形成するので、表面圧を調整
することなく、自動的に均一な担持液膜を形成し、液液
界面における単分子膜を形成することができるので、均
一な単分子膜の作成が容易で、蛋白２次元結晶膜の分解
能を著しく向上させることができる。(Effects of the Invention) As is clear from the above description, according to the present invention, the anchor film represented by the water surface film is formed by a liquid film having excellent lateral flowability without using a surfactant. A uniform supported liquid film can be automatically formed without adjusting the surface pressure, and a monolayer at the liquid-liquid interface can be formed. The resolution of the film can be significantly improved.
尚、本発明は上記実施例に基づいて説明されたが、通
常の表面膜作成トラフを用いても作成することができ、
その場合、アンカー膜となる液膜は仕切り部材により表
面圧を調整する必要がない。Although the present invention has been described based on the above embodiment, the present invention can also be formed using a normal surface film forming trough,
In this case, it is not necessary to adjust the surface pressure of the liquid film serving as the anchor film by the partition member.
第１図は本発明方法を適用するに適する単分子膜作成装
置の概要図、第２図は従来の単分子膜作成の１例を示す
概念図である。 １……トラフ、11……収容部FIG. 1 is a schematic diagram of a monolayer forming apparatus suitable for applying the method of the present invention, and FIG. 2 is a conceptual diagram showing an example of a conventional monolayer forming. 1 ... trough, 11 ... accommodation section
Claims (2)
膜形成室内に入れ、該蛋白質水溶液面に、蛋白分子に対
し吸着性を示す第１級アミノ基を有する水に不溶な液体
の層からなる液膜を形成し、該蛋白質水溶液面と該液膜
との液液界面において、該液膜を担持液膜として、該液
膜に蛋白分子を吸着・担持させる工程を含む蛋白２次元
結晶膜作成方法。1. An aqueous protein solution is placed in a water-repellent recess or a film formation chamber in a trough, and a water-insoluble liquid layer having a primary amino group having an adsorptivity to protein molecules is formed on the surface of the aqueous protein solution. A two-dimensional crystal film comprising the steps of: forming a liquid film, and adsorbing and supporting protein molecules on the liquid film at the liquid-liquid interface between the aqueous protein solution surface and the liquid film, using the liquid film as a supporting liquid film. How to make.
その誘導体からなる請求項１記載の蛋白２次元結晶膜作
成方法。2. The method according to claim 1, wherein the liquid film is made of dehydroabiethylamine or a derivative thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18804989A JP2895097B2 (en) | 1989-07-20 | 1989-07-20 | Method for preparing protein two-dimensional crystal film |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18804989A JP2895097B2 (en) | 1989-07-20 | 1989-07-20 | Method for preparing protein two-dimensional crystal film |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0352897A JPH0352897A (en) | 1991-03-07 |
| JP2895097B2 true JP2895097B2 (en) | 1999-05-24 |
Family
ID=16216791
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP18804989A Expired - Lifetime JP2895097B2 (en) | 1989-07-20 | 1989-07-20 | Method for preparing protein two-dimensional crystal film |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2895097B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SG189159A1 (en) * | 2010-10-27 | 2013-05-31 | Sigma Tau Ind Farmaceuti | Diterpenoid derivatives endowed of biological properties |
-
1989
- 1989-07-20 JP JP18804989A patent/JP2895097B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0352897A (en) | 1991-03-07 |
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