Jabeen Talat, Sharma Sujata, Singh Nagendra, Singh Rajendra K, Verma Ashok K, Paramasivam M, Srinivasan A, Singh Tej P
Department of Biophysics, All India Institute of Medical Sciences, New Delhi 110029, India.
Acta Crystallogr D Biol Crystallogr. 2005 Mar;61(Pt 3):302-8. doi: 10.1107/S0907444904033165. Epub 2005 Feb 24.
The crystal structure of a zinc-induced heterodimer of two metal-free isoforms of a cobra venom phospholipase A(2) has been determined at 2.7 angstroms resolution. The crystals belong to space group P4(1), with unit-cell parameters a = b = 65.5, c = 58.4 angstroms, and have a single dimer in the asymmetric unit. The structure has been refined to R(cryst) and R(free) factors of 0.188 and 0.232, respectively. The two isoforms have a sequence identity of 82%. The zinc ion forms a fivefold coordination with a trigonal bipyramidal geometry involving one O atom each from Asp24 and Asn112 from molecule A and Asp24 from molecule B and two water molecules. Both molecules of the dimer are inactive. Molecule A is inactive because Arg31 (B) binds to Asp49 (A), while an acetate ion has displaced the essential water molecule and interacts with His48 (A). On the other hand, Arg31 (A) interacts with the calcium-binding loop of molecule B, resulting in an altered conformation of the loop. The absence of a calcium ion, loss of the essential water molecule and the altered conformation of the calcium-binding loop may be the reasons for the loss of activity of molecule B.
已在2.7埃分辨率下测定了锌诱导的眼镜蛇毒磷脂酶A(2)两种无金属同工型的异二聚体的晶体结构。晶体属于空间群P4(1),晶胞参数a = b = 65.5,c = 58.4埃,不对称单元中有一个二聚体。该结构已精修至R(cryst)和R(free)因子分别为0.188和0.232。这两种同工型的序列同一性为82%。锌离子形成五配位,呈三角双锥几何构型,涉及来自分子A的Asp24和Asn112各一个O原子、来自分子B的Asp24以及两个水分子。二聚体的两个分子均无活性。分子A无活性是因为Arg31 (B)与Asp49 (A)结合,而一个醋酸根离子取代了必需的水分子并与His48 (A)相互作用。另一方面,Arg31 (A)与分子B的钙结合环相互作用,导致该环的构象改变。钙离子的缺失、必需水分子的丧失以及钙结合环构象的改变可能是分子B失去活性的原因。