Literature summary for 5.1.1.5 extracted from

Tape, C.J.; Norrie, I.C.; Worboys, J.D.; Lim, L.; Lauffenburger, D.A.; Joergensen, C.
Cell-specific labeling enzymes for analysis of cell-cell communication in continuous co-culture (2014), Mol. Cell. Proteomics, 13, 1866-1876 .

Application

Application	Comment	Organism
additional information	the enzyme is used for isotopic labeling of cells	Proteus mirabilis

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of C-terminally HA-tagged wild-type enzyme and mutant enzyme LyrM37-KDEL, codon optimized for mouse expression, in C3H10T1/2 cells or MDA-MB-231 cells. MDA-MB-231 cells are infected with pGIPZ lentivirus for GFP expression, and C3H10T1/2 cells are infected with pMSCV-pBabeMCS-IRES-RFP retrovirus for RFP expression, identification of proteotypic Lyr peptides suitable for relative isotopic quantification. Wild-type Proteus mirabilis Lyr is prolifically secreted from eukaryotic cells in contrast to mutant enzyme LyrM37-KDEL. Extracellular Lyr converts labeled D-lysine to labeled L-lysine in conditioned media and severely compromises coculture labeling efficiency. Cells stably transfected with LyrM37-KDEL achieve proliferation comparable to that with L-lysine when grown on concentrations of D-lysine greater than 1 mM	Proteus mirabilis

Cloned (Comment)

Organism

recombinant expression of C-terminally HA-tagged wild-type enzyme and mutant enzyme LyrM37-KDEL, codon optimized for mouse expression, in C3H10T1/2 cells or MDA-MB-231 cells. MDA-MB-231 cells are infected with pGIPZ lentivirus for GFP expression, and C3H10T1/2 cells are infected with pMSCV-pBabeMCS-IRES-RFP retrovirus for RFP expression, identification of proteotypic Lyr peptides suitable for relative isotopic quantification. Wild-type Proteus mirabilis Lyr is prolifically secreted from eukaryotic cells in contrast to mutant enzyme LyrM37-KDEL. Extracellular Lyr converts labeled D-lysine to labeled L-lysine in conditioned media and severely compromises coculture labeling efficiency. Cells stably transfected with LyrM37-KDEL achieve proliferation comparable to that with L-lysine when grown on concentrations of D-lysine greater than 1 mM

Proteus mirabilis

Protein Variants

Protein Variants	Comment	Organism
additional information	in an attempt to limit extracellular Lyr (while retaining the catalytic activity), amino acids 1-36 are removed from the enzyme (LyrM37) and a C-terminal KDEL ER retention motif (LyrM37-KDEL) is added	Proteus mirabilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-lysine	Proteus mirabilis	-	D-lysine	-	r

Organism

Organism	UniProt	Comment	Textmining
Proteus mirabilis	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-lysine	-	Proteus mirabilis	D-lysine	-	r

Subunits

Subunits	Comment	Organism
More	Lyr contains a globular catalytic core (amino acids 37-407) distinct from the putative signal peptide	Proteus mirabilis

Synonyms

Synonyms	Comment	Organism
lyr	-	Proteus mirabilis

General Information

General Information	Comment	Organism
additional information	unlike wild-type enzyme LyrWT, enzyme mutant LyrM37-KDEL is a truly intracellular D-lysine conversion enzyme	Proteus mirabilis
physiological function	enzyme Lyr catalyzes the conversion of D-lysine into L-lysine. Proteus mirabilis Lyr activity is independent of its putative signal peptide and can function in the eukaryotic endoplasmic reticulum	Proteus mirabilis