Expression and crystallization of several forms of the Propionibacterium shermanii transcarboxylase 5S subunit.
The dimeric outer 5S subunit of transcarboxylase has been expressed in three different forms and crystallized: native 5S, 5S-His(6) and selenomethione-5S-His(6). All the crystals have an orthorhombic space group, but while native 5S forms primitive orthorhombic crystals, 5S-His(6) crystals are either C-centered or primitive and SeMet-5S-His(6) crystals are C-centered. Crystallization of native 5S requires the addition of lithium sulfate, whereas this salt prevented crystallization of 5S-His(6). All 5S crystals diffract to approximately 2.0 A resolution with synchrotron radiation. Efforts are under way to solve the structure of SeMet-5S-His(6) using MAD.
Department of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
Pamela R
PR
Hall
Run
R
Zheng
Marianne
M
Pusztai-Carey
Focco
F
van den Akker
Paul R
PR
Carey
Vivien C
VC
Yee
D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D019878:Carboxyl and Carbamoyl Transferases / Q000737:chemistry* / Q000302:isolation & purification*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006639:Histidine / Q000737:chemistry; D018020:Lithium Compounds / Q000737:chemistry; D011424:Propionibacterium / Q000201:enzymology*; D021122:Protein Subunits / Q000737:chemistry* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012645:Selenomethionine / Q000737:chemistry
D001426:Bacterial Proteins; D018020:Lithium Compounds; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D006639:Histidine; D012645:Selenomethionine; D019878:Carboxyl and Carbamoyl Transferases; C021580:Methylmalonyl-CoA carboxytransferase