Samson I, Kerremans L, Rozenski J, Samyn B, Van Beeumen J, Van Aerschot A, Herdewijn P
Laboratory of Medicinal Chemistry (F.F.W.), Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium.
Bioorg Med Chem. 1995 Mar;3(3):257-65. doi: 10.1016/0968-0896(95)00020-h.
A synthetic peptide library, composed of 2.5 million L-amino acid pentapeptides anchored on polystyrene beads was prepared with each bead bearing a single pentapeptide sequence. This library was screened for interaction with glycosomal phosphoglycerate kinase (gPGK) of Trypanosoma brucei labelled with fluorescein or with biotin. Affinity beads that bound the enzyme were selected with a pipette or with streptavidin coated magnetic beads. The beads that bound to the enzyme were individually subjected to Edman microsequence analysis to determine the sequence of the corresponding peptide ligands. The corresponding peptide-sequences were synthesised as free peptide acids and evaluated for enzyme activity inhibition. The pentapeptide NWMMF was able to selectively inhibit gPGK with an IC50 of approximately 80 microM.
制备了一个合成肽库，由固定在聚苯乙烯珠上的250万个L - 氨基酸五肽组成，每个珠子带有一个单一的五肽序列。用荧光素或生物素标记布氏锥虫的糖体磷酸甘油酸激酶（gPGK），对该肽库进行与gPGK相互作用的筛选。用移液器或链霉亲和素包被的磁珠选择与酶结合的亲和珠。将与酶结合的珠子分别进行埃德曼微量序列分析，以确定相应肽配体的序列。将相应的肽序列合成为游离肽酸，并评估其对酶活性的抑制作用。五肽NWMMF能够选择性抑制gPGK，IC50约为80微摩尔。
