Methods of detecting and measuring glutamine and analogues thereof, and methods related thereto
Method for the detection of glutamine and its analogs are provided by the present disclosure. Also provided are methods for measuring the levels of glutamine and its analogs, including diagnostic methods. Further provided are methods that utilize glutamine analogs for the synthesis of colored pigments and other useful agents. The methods comprise the use of a nonribosomal peptide synthetase (NRPS) under conditions to produce an indigoidine or indigoidine-related pigment.
1. A method of diagnosing in a test subject a condition or disorder associated with altered levels of L-glutamine or an L-glutamine analogue, by measuring the concentration of L-glutamine or an L-glutamine analogue in a sample from the subject, comprising:
a) incubating the sample with an activated pigment-producing non-ribosomal peptide synthase (NRPS) to produce indigoidine pigment or a derivatized indigoidine pigment; and
b) quantifying the amount of the indigoidine pigment or the derivatized indigoidine pigment produced, thereby measuring the concentration of the L-glutamine or the L-glutamine analogue, respectively, in the sample.
2. The method of claim 1 , wherein the pigment-producing NRPS is blue pigment synthetase A (BpsA), wherein the BpsA is activated prior to isolation or following isolation of the BpsA.
3. The method of claim 2 , wherein the sample is a biological sample selected from the group consisting of a blood sample, a urine sample, a saliva sample, a cerebrospinal fluid sample, a lymph fluid sample, and a eukaryotic cell culture sample.
4. The method of claim 2 , wherein the L-glutamine analogue is selected from the group consisting of glycyl-glutamine, N-trifluoroacetyl-L-glutamine, alanyl-glutamine, indoleacetyl glutamine, N-acetylglutamine, phenylacetyl-L-glutamine, and glutamine-terminal peptides or polypeptides.
5. The method of claim 1 ,
wherein an altered amount of the indigoidine pigment or the derivatized indigoidine pigment relative to a control or standard amount indicates altered levels of the L-glutamine or the L-glutamine analogue, respectively.
6. The method of claim 1 , wherein the condition or disorder associated with altered levels of L-glutamine is selected from the group consisting of overtraining syndrome, urea cycle disorders, Alzheimer's disease and other neurodegenerative disorders, cancers, and aminoaciduria.
7. The method of claim 1 , wherein the glutamine analogue is selected from the group consisting of indoleacetyl glutamine, N-acetylglutamine, and phenylacetyl-L-glutamine.
8. The method of claim 1 , wherein the condition or disorder associated with altered levels of the L-glutamine analogue is selected from the group consisting of Hartnup disease, autosomal dominant polycystic kidney disease, renal tubal injury, aminoacylase deficiency, and uremia.