TY - JOUR
T1 - The structural basis of chain length control in Rv1086
AU - Wang, Wenjian
AU - Dong, Changjiang
AU - McNeil, Michael
AU - Kaur, Devinder
AU - Mahapatra, Sebabrata
AU - Crick, Dean C.
AU - Naismith, James H.
PY - 2008/8/1
Y1 - 2008/8/1
N2 - In Mycobacterium tuberculosis, two related Z-prenyl diphosphate synthases, E,Z-farnesyl diphosphate synthase (Rv1086) and decaprenyl diphosphate synthase (Rv2361c), work in series to synthesize decaprenyl phosphate (C,50) from isopentenyl diphosphate and E-geranyl diphosphate. Decaprenyl phosphate plays a central role in the biosynthesis of essential mycobacterial cell wall components, such as the mycolyl-arabinogalactan-peptidoglycan complex and lipoarabinomannan; thus, its synthesis has attracted considerable interest as a potential therapeutic target. Rv1086 is a unique prenyl diphosphate synthase in that it adds only one isoprene unit to geranyl diphosphate, generating the 15-carbon product (E,Z-famesyl diphosphate). Rv2361c then adds a further seven isoprene units to E,Z-farnesyl diphosphate in a processive manner to generate the 50-carbon prenyl diphosphate, which is then dephosphorylated to generate a carrier for activated sugars. The molecular basis for chain-length discrimination by Rv1086 during synthesis is unknown. We also report the structure of apo Rv1086 with citronellyl diphosphate bound and with the product mimic E,E-famesyl diphosphate bound. We report the structures of Rv2361c in the apo form, with isopentenyl diphosphate bound and with a substrate analogue, citronellyl diphosphate. The structures confirm the enzymes are very closely related. Detailed comparison reveals structural differences that account for chain-length control in Rv1086. We have tested this hypothesis and have identified a double mutant of Rv1086 that makes a range of longer lipid chains. (C) 2008 Elsevier Ltd. All rights reserved.
AB - In Mycobacterium tuberculosis, two related Z-prenyl diphosphate synthases, E,Z-farnesyl diphosphate synthase (Rv1086) and decaprenyl diphosphate synthase (Rv2361c), work in series to synthesize decaprenyl phosphate (C,50) from isopentenyl diphosphate and E-geranyl diphosphate. Decaprenyl phosphate plays a central role in the biosynthesis of essential mycobacterial cell wall components, such as the mycolyl-arabinogalactan-peptidoglycan complex and lipoarabinomannan; thus, its synthesis has attracted considerable interest as a potential therapeutic target. Rv1086 is a unique prenyl diphosphate synthase in that it adds only one isoprene unit to geranyl diphosphate, generating the 15-carbon product (E,Z-famesyl diphosphate). Rv2361c then adds a further seven isoprene units to E,Z-farnesyl diphosphate in a processive manner to generate the 50-carbon prenyl diphosphate, which is then dephosphorylated to generate a carrier for activated sugars. The molecular basis for chain-length discrimination by Rv1086 during synthesis is unknown. We also report the structure of apo Rv1086 with citronellyl diphosphate bound and with the product mimic E,E-famesyl diphosphate bound. We report the structures of Rv2361c in the apo form, with isopentenyl diphosphate bound and with a substrate analogue, citronellyl diphosphate. The structures confirm the enzymes are very closely related. Detailed comparison reveals structural differences that account for chain-length control in Rv1086. We have tested this hypothesis and have identified a double mutant of Rv1086 that makes a range of longer lipid chains. (C) 2008 Elsevier Ltd. All rights reserved.
KW - drug design
KW - enzyme mechanism
KW - tuberculosis
KW - x-ray crystallography
KW - inhibitors
KW - UNDECAPRENYL DIPHOSPHATE SYNTHASE
KW - MYCOBACTERIUM-TUBERCULOSIS
KW - BIOSYNTHESIS
KW - COMPLEXES
UR - http://www.scopus.com/inward/record.url?scp=46649099251&partnerID=8YFLogxK
UR - http://ukpmc.ac.uk/abstract/MED/18597781
U2 - 10.1016/j.jmb.2008.05.060
DO - 10.1016/j.jmb.2008.05.060
M3 - Article
SN - 0022-2836
VL - 381
SP - 129
EP - 140
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 1
ER -