Andersson B, Anderson J M, Ryrie I J
Eur J Biochem. 1982 Apr 1;123(2):465-72. doi: 10.1111/j.1432-1033.1982.tb19790.x.
To investigate the transverse bilayer organization of the chlorophyll-proteins of the three intrinsic chlorophyll-protein complexes, the effects of proteolytic enzymes, and an antibody against the light-harvesting complex were compared using right-side-out and inside-out thylakoid vesicles. The vesicles were isolated by aqueous polymer phase partitioning following the fragmentation of spinach thylakoids by passage through a Yeda press. Both vesicle types were agglutinated by an antiserum specific for the light-harvesting complex, although proteolytic degradation of the complex occurred only in right-side-out vesicles. In addition, there are different antigenic sites for the light-harvesting complex on the inner and outer thylakoid surfaces. Polypeptides of the chlorophyll-alpha-protein complex of photosystem II were degraded by proteases at both membrane surfaces. We concluded that both these chlorophyll-protein complexes are membrane spanning and transversely asymmetric, but that the light-harvesting complex polypeptides accessible at the inner thylakoid surface are more resistant to proteolytic attack. In contrast, the main chlorophyll-containing polypeptide (Mr = 64 500) of photosystem I complex was resistant to proteolytic attack at both the outer and inner thylakoid surfaces.
为了研究三种内在叶绿素 - 蛋白质复合物中叶绿素 - 蛋白质的横向双层组织，使用外翻和内翻类囊体囊泡比较了蛋白水解酶和抗捕光复合物抗体的作用。通过Yeda压榨机破碎菠菜类囊体后，采用水相聚合物相分配法分离囊泡。两种类型的囊泡都能被抗捕光复合物的抗血清凝集，尽管该复合物的蛋白水解降解仅发生在外翻囊泡中。此外，类囊体内外表面上的捕光复合物存在不同的抗原位点。光系统II的叶绿素 - α - 蛋白质复合物的多肽在两个膜表面都被蛋白酶降解。我们得出结论，这两种叶绿素 - 蛋白质复合物都是跨膜的且横向不对称，但类囊体内表面可及的捕光复合物多肽对蛋白水解攻击更具抗性。相比之下，光系统I复合物的主要含叶绿素多肽（Mr = 64500）在类囊体外表面和内表面都对蛋白水解攻击具有抗性。
