Structural and functional studies of fatty acyl adenylate ligases from E. coli and L. pneumophila.
Structural and functional studies of fatty acyl adenylate ligases from E. coli and L. pneumophila.
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DOI：
10.1016/j.jmb.2010.12.011
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发表时间：
2011-02-18
影响因子：
5.6
通讯作者：
Swaminathan S
中科院分区：
文献类型：
--
作者：
Zhang Z;Zhou R;Sauder JM;Tonge PJ;Burley SK;Swaminathan S
脂肪酰基腺苷酸连接酶（FAAL）是近年来在结核分枝杆菌（Mtb）中发现的腺苷酸形成酶家族的新成员。它们在序列上与脂肪酰辅酶A连接酶（FACL）相似。然而，虽然FACL进行两步催化反应，AMP连接，然后使用ATP和CoA作为辅因子进行CoA连接，但FAAL仅产生酰基腺苷酸，并且不能进行第二步。本文报道了E.大肠杆菌（EcFAAL）和嗜肺军团菌（LpFAAL）与酰基腺苷酸结合，分别在3.0和1.85 μ M的分离限下测定。这些结构共有一个较大的N-末端结构域和一个较小的C-末端结构域，它们共同类似于先前确定的FAAL和FACL蛋白的结构。我们的两种结构发生在完全不同的构象。EcFAAL采用FACL典型的腺苷酸形成构象，而LpFAAL表现出独特的中间构象。EcFAAL和LpFAAL都具有将它们与FACL区分开的插入基序。EcFAAL和LpFAAL的结构揭示了该插入基序与域间铰链区以及与C-末端结构域之间的详细相互作用。我们建议，插入基序支持足够的域间运动，使底物结合和产品释放过程中酰基腺苷酸的形成，而他们排除CoA结合，从而防止CoA连接。
Fatty acyl-AMP ligase (FAAL) is a new member of a family of adenylate-forming enzymes that were recently discovered in Mycobacterium tuberculosis (Mtb). They are similar in sequence to fatty acyl-CoA ligases (FACLs). However, while FACLs perform a two-step catalytic reaction, AMP ligation followed by CoA ligation using ATP and CoA as cofactors, FAALs produce only the acyl adenylate and are unable to perform the second step. We report X-ray crystal structures of full length FAALs from E. coli (EcFAAL) and Legionella pneumophila (LpFAAL) bound to acyl adenylate, determined at resolution limits of 3.0 and 1.85 Å, respectively. The structures share a larger N-terminal domain and a smaller C-terminal domain, which together resemble the previously determined structures of FAAL and FACL proteins. Our two structures occur in quite different conformations. EcFAAL adopts the adenylate forming conformation typical of FACLs, whereas LpFAAL exhibits a unique intermediate conformation. Both EcFAAL and LpFAAL have insertion motifs that distinguish them from the FACLs. Structures of EcFAAL and LpFAAL reveal detailed interactions between this insertion motif and the interdomain hinge region and with the C-terminal domain. We suggest that the insertion motifs support sufficient interdomain motions to allow substrate binding and product release during acyl adenylate formation, whereas they preclude CoA binding thereby preventing CoA ligation.
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影响因子：
14.8
作者：
Arora, Pooja;Goyal, Aneesh;Natarajan, Vivek T.;Rajakumara, Eerappa;Verma, Priyanka;Gupta, Radhika;Yousuf, Malikmohamed;Trivedi, Omita A.;Mohanty, Debasisa;Tyagi, Anil;Sankaranarayanan, Rajan;Gokhale, Rajesh S.
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Gokhale, Rajesh S.
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
4.8
作者：
Black, PN;Zhang, Q;DiRusso, CC
通讯作者：
DiRusso, CC
影响因子：
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作者：
Leger, Mathieu;Gavalda, Sabine;Marrakchi, Hedia
通讯作者：
Marrakchi, Hedia
DOI：
10.1107/s0907444902016657
发表时间：
2002-11-01
影响因子：
2.2
作者：
Adams, PD;Grosse-Kunstleve, RW;Terwilliger, TC
通讯作者：
Terwilliger, TC