5HPG

STRUCTURE AND LIGAND DETERMINANTS OF THE RECOMBINANT KRINGLE 5 DOMAIN OF HUMAN PLASMINOGEN

5HPG の概要

• エントリーDOI: 10.2210/pdb5hpg/pdb
• 分子名称: PLASMINOGEN (2 entities in total)
• 機能のキーワード: serine protease, kringle 5, human plasminogen, fibrinolysis
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Secreted: P00747
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 18788.88

構造登録者

Tulinsky, A.,Mochalkin, I.,Castellino, F.J. (登録日: 1997-09-19, 公開日: 1998-03-25, 最終更新日: 2024-10-23)

主引用文献

Chang, Y.,Mochalkin, I.,McCance, S.G.,Cheng, B.,Tulinsky, A.,Castellino, F.J.
Structure and ligand binding determinants of the recombinant kringle 5 domain of human plasminogen.
Biochemistry, 37:3258-3271, 1998

PubMed Abstract: The X-ray crystal structure of the recombinant (r) kringle 5 domain of human plasminogen (K5HPg) has been solved by molecular replacement methods using K1HPg as a model and refined at 1.7 A resolution to an R factor of 16.6%. The asymmetric unit of K5HPg is composed of two molecules related by a noncrystallographic 2-fold rotation axis approximately parallel to the z-direction. The lysine binding site (LBS) is defined by the regions His33-Thr37, Pro54-Val58, Pro61-Tyr64, and Leu71-Tyr74 and is occupied in the apo-form by water molecules. A unique feature of the LBS of apo-K5HPg is the substitution by Leu71 for the basic amino acid, arginine, that in other kringle polypeptides forms the donor cationic center for the carboxylate group of omega-amino acid ligands. While wild-type (wt) r-K5HPg interacted weakly with these types of ligands, replacement by site-directed mutagenesis of Leu71 by arginine led to substantially increased affinity of the ligands for the LBS of K5HPg. As a result, binding of omega-amino acids to this mutant kringle (r-K5HPg[L71R]) was restored to levels displayed by the companion much stronger affinity HPg kringles, K1HPg and K4HPg. Correspondingly, alkylamine binding to r-K5HPg[L71R] was considerably attenuated from that shown by wtr-K5HPg. Thus, employing a rational design strategy based on the crystal structure of K5HPg, successful remodeling of the LBS has been accomplished, and has resulted in the conversion of a weak ligand binding kringle to one that possesses an affinity for omega-amino acids that is similar to K1HPg and K4HPg.

PubMed: 9521645
DOI: 10.1021/bi972284e

実験手法

X-RAY DIFFRACTION (1.66 Å)