Structural and Biochemical Insight into the Recruitment of Acyl Carrier Protein‐Linked Extender Units in Ansamitocin Biosynthesis

ChemBioChem ◽  
2020 ◽  
Vol 21 (9) ◽  
pp. 1309-1314 ◽  
Author(s):  
Fa Zhang ◽  
Huining Ji ◽  
Imtiaz Ali ◽  
Zixin Deng ◽  
Linquan Bai ◽  
...  
Keyword(s):  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
Insight Into

scholarly journals Mycobacterium tuberculosis Proteasome Accessory Factor A (PafA) Can Transfer Prokaryotic Ubiquitin-Like Protein (Pup) between Substrates

mBio ◽  
2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Susan Zhang ◽  
Kristin E. Burns-Huang ◽  
Guido V. Janssen ◽  
Huilin Li ◽  
Huib Ovaa ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Protein Stability ◽  
Protein Degradation ◽  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
Inositol 1 ◽  
Accessory Factor ◽  
Alternative Sources ◽  
Insight Into

ABSTRACT The protein degradation machinery of Mycobacterium tuberculosis includes a proteasome and a ubiquitin-like protein (Pup). Proteasome accessory factor A (PafA) attaches Pup to proteins to target them for degradation by the proteasome. Free Pup is unstable and never observed in extracts of M. tuberculosis , an observation that led us to hypothesize that PafA may need alternative sources of Pup. Here, we show that PafA can move Pup from one proteasome substrate, inositol 1-phosphate synthetase (Ino1), to two different proteins, malonyl coenzyme A (CoA)-acyl carrier protein transacylase (FabD) and lonely guy (Log). This apparent “transpupylation” reaction required a previously unrecognized depupylase activity in PafA, and, surprisingly, this depupylase activity was much more efficient than the activity of the dedicated depupylase Dop (deamidase of Pup). Thus, PafA can potentially use both newly synthesized Pup and recycled Pup to doom proteins for degradation. IMPORTANCE Unlike eukaryotes, which contain hundreds of ubiquitin ligases, Pup-containing bacteria appear to have a single ligase to pupylate dozens if not hundreds of different proteins. The observation that PafA can depupylate and transpupylate in vitro offers new insight into how protein stability is regulated in proteasome-bearing bacteria. Importantly, PafA and the dedicated depupylase Dop are each required for the full virulence of Mycobacterium tuberculosis . Thus, inhibition of both enzymes may be extremely attractive for the development of therapeutics against tuberculosis.

scholarly journals Identification of the Binding Region of the [2Fe-2S] Ferredoxin in Stearoyl-Acyl Carrier Protein Desaturase:  Insight into the Catalytic Complex and Mechanism of Action†

Biochemistry ◽  
2006 ◽  
Vol 45 (15) ◽  
pp. 4848-4858 ◽  
Author(s):  
Pablo Sobrado ◽  
Karen S. Lyle ◽  
Steven P. Kaul ◽  
Michelle M. Turco ◽  
Ida Arabshahi ◽  
...  
Keyword(s):  
Mechanism Of Action ◽  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
Catalytic Complex ◽  
Binding Region ◽  
Insight Into

scholarly journals Solution Structures of Conformationally Equilibrium Forms of Holo-Acyl Carrier Protein (PfACP) fromPlasmodium falciparumProvides Insight into the Mechanism of Activation of ACPs†,‡

Biochemistry ◽  
2006 ◽  
Vol 45 (22) ◽  
pp. 6904-6916 ◽  
Author(s):  
Alok Kumar Sharma ◽  
Shailendra Kumar Sharma ◽  
Avadhesha Surolia ◽  
Namita Surolia ◽  
Siddhartha P. Sarma
Keyword(s):  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
Solution Structures ◽  
Insight Into

scholarly journals Structure-based analysis of the molecular interactions between acyltransferase and acyl carrier protein in vicenistatin biosynthesis

2016 ◽  
Vol 113 (7) ◽  
pp. 1802-1807 ◽  
Author(s):  
Akimasa Miyanaga ◽  
Shohei Iwasawa ◽  
Yuji Shinohara ◽  
Fumitaka Kudo ◽  
Tadashi Eguchi
Keyword(s):  
Crystal Structure ◽  
Protein Interactions ◽  
Acyl Carrier Protein ◽  
Complex Structure ◽  
Binding Pocket ◽  
Carrier Protein ◽  
Protein Protein Interactions ◽  
Substrate Binding Pocket ◽  
Insight Into

Acyltransferases (ATs) are key determinants of building block specificity in polyketide biosynthesis. Despite the importance of protein–protein interactions between AT and acyl carrier protein (ACP) during the acyltransfer reaction, the mechanism of ACP recognition by AT is not understood in detail. Herein, we report the crystal structure of AT VinK, which transfers a dipeptide group between two ACPs, VinL and VinP1LdACP, in vicenistatin biosynthesis. The isolated VinK structure showed a unique substrate-binding pocket for the dipeptide group linked to ACP. To gain greater insight into the mechanism of ACP recognition, we attempted to crystallize the VinK–ACP complexes. Because transient enzyme–ACP complexes are difficult to crystallize, we developed a covalent cross-linking strategy using a bifunctional maleimide reagent to trap the VinK–ACP complexes, allowing the determination of the crystal structure of the VinK–VinL complex. In the complex structure, Arg-153, Met-206, and Arg-299 of VinK interact with the negatively charged helix II region of VinL. The VinK–VinL complex structure allows, to our knowledge, the first visualization of the interaction between AT and ACP and provides detailed mechanistic insights into ACP recognition by AT.

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