scholarly journals Resin supported acyl carrier protein labeling strategies

RSC Advances ◽  
2014 ◽  
Vol 4 (18) ◽  
pp. 9092-9097 ◽  
Author(s):  
Michael Rothmann ◽  
Nicolas M. Kosa ◽  
Michael D. Burkart
Keyword(s):  
Acyl Carrier Protein ◽  
Protein Labeling ◽  
Carrier Protein ◽  
Carrier Proteins ◽  
Acyl Carrier Proteins ◽  
Functional Modification ◽  
Labeling Strategies

The post-translational modifying enzymes phophopantetheinyl transferase and acyl carrier protein hydrolase show utility as resin supported conjugates in the functional modification of acyl carrier proteins.

scholarly journals Products of fatty acid synthesis by a particulate fraction from germinating pea (Pisum sativum L.)

1981 ◽  
Vol 199 (1) ◽  
pp. 221-226 ◽  
Author(s):  
J Sanchez ◽  
J L Harwood
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Pisum Sativum ◽  
Acyl Carrier Protein ◽  
Acid Synthesis ◽  
Simultaneous Increase ◽  
Carrier Protein ◽  
Carrier Proteins ◽  
Acyl Carrier Proteins ◽  
Malonyl Coa

The synthesis of lipids and acyl thioesters was studied in microsomal preparations from germinating pea (Pisum sativum cv. Feltham First) seeds. Under conditions of maximal synthesis (in the presence of exogenous acyl-carrier protein) acyl-acyl-carrier proteins accounted for about half the total incorporation from [14C]malonyl-CoA. Decreasing the concentrations of exogenous acyl-carrier protein lowered the overall synthesis of fatty acids by decreasing, almost exclusively, the radioactivity associated with acyl-acyl-carrier proteins. A time-course experiment showed that acyl-acyl-carrier proteins accumulated most of the radioactive label at the beginning of the incubation but, eventually, the amount of radioactivity in that fraction decreased, while a simultaneous increase in the acyl-CoA and lipid fractions was noticed. Addition of exogenous CoA (1 mM) produced a decrease of total incorporation, but an increase in the radioactivity incorporated into acyl-CoA. The microsomal preparations synthesized saturated fatty acids up to C20, including significant proportions of pentadecanoic acid and heptadecanoic acid. Synthesis of these ‘odd-chain’ fatty acids only took place in the microsomal fraction. In contrast, when the 18,000g supernatant (containing the microsomal and soluble fractions) was incubated with [14C]malonyl-CoA, the radioactive fatty acid and acyl classes closely resembled the patterns produced by germinating in the presence of [14C]acetate in vivo. The results are discussed in relation to the role of acyl thioesters in the biosynthesis of plant lipids.

scholarly journals Purification and separation of holo- and apo-forms of Saccharopolyspora erythraea acyl-carrier protein released from recombinant Escherichia coli by freezing and thawing

1993 ◽  
Vol 294 (2) ◽  
pp. 521-527 ◽  
Author(s):  
S A Morris ◽  
W P Revill ◽  
J Staunton ◽  
P F Leadlay
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Acyl Carrier Protein ◽  
Saccharopolyspora Erythraea ◽  
Carrier Protein ◽  
Freezing And Thawing ◽  
Acyl Carrier Proteins ◽  
Expression Plasmid ◽  
E Coli ◽  
Protein Dimer

Saccharopolyspora erythraea acyl-carrier protein, highly expressed from a T7-based expression plasmid in Escherichia coli, can be selectively released from the cells in near-quantitative yield by a single cycle of freezing and thawing in a neutral buffer. Electrospray mass spectrometry was used to confirm that the recombinant S. erythraea acyl-carrier protein over-expressed in E. coli is present predominantly as the holo-form, with variable amounts of apo-acyl-carrier protein, holo-acyl-carrier protein dimer and holo-acyl-carrier protein glutathione adduct. The holo- and apo-acyl-carrier proteins are both readily purified on a large scale from the freeze-thaw extracts and can be separated from one another by octyl-Sepharose chromatography. The holo-acyl-carrier protein obtained in this way was fully active in supporting the synthesis of acyl-acyl-carrier protein by extracts of S. erythraea.

scholarly journals Rv0100, a proposed acyl carrier protein in Mycobacterium tuberculosis: expression, purification and crystallization

2019 ◽  
Vol 75 (10) ◽  
pp. 646-651 ◽  
Author(s):  
Jasper Marc G. Bondoc ◽  
Hiten J. Gutka ◽  
Mashal M. Almutairi ◽  
Ryan Patwell ◽  
Maxwell W. Rutter ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Smegmatis ◽  
Drug Target ◽  
Fatty Acid Biosynthesis ◽  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
Acid Biosynthesis ◽  
Acyl Carrier Proteins ◽  
Drug Discovery And Development ◽  
Novel Drug

Acyl carrier proteins (ACPs) are important components in fatty-acid biosynthesis in prokaryotes. Rv0100 is predicted to be an essential ACP in Mycobacterium tuberculosis, the pathogen that is the causative agent of tuberculosis, and therefore has the potential to be a novel antituberculosis drug target. Here, the successful cloning and purification of Rv0100 using Mycobacterium smegmatis as a host is reported. Crystals of the purified protein were obtained that diffracted to a resolution of 1.9 Å. Overall, this work lays the foundation for the future pursuit of drug discovery and development against this potentially novel drug target.

scholarly journals N-Activated β-lactams as versatile reagents for acyl carrier protein labeling

2012 ◽  
Vol 10 (10) ◽  
pp. 1992 ◽  
Author(s):  
Gitanjeli Prasad ◽  
Jon W. Amoroso ◽  
Lawrence S. Borketey ◽  
Nathan A. Schnarr
Keyword(s):  
Acyl Carrier Protein ◽  
Protein Labeling ◽  
Carrier Protein
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