Introduction
Pyruvate kinase (PKLR) is an essential enzyme that plays a critical role in glycolysis, the metabolic pathway responsible for breaking down glucose to produce energy. It catalyzes the final step of glycolysis, converting phosphoenolpyruvate (PEP) to pyruvate, a crucial molecule in cellular energy production. PKLR exists in different isoforms, including L- and R-types, encoded by the PKLR gene. The L-type isoform is particularly relevant in glucose metabolism and has been linked to maturity-onset diabetes of the young.
Description
This product is a recombinant human PKLR protein expressed in E. coli and purified to a high degree. It is a single, non-glycosylated polypeptide chain with a molecular weight of 59.2 kDa. The protein sequence comprises amino acids 47 to 574 of the native human PKLR, with a 21-amino acid His-tag fused at the N-terminus to facilitate purification.
Physical Appearance
The product appears as a clear, colorless solution that has been sterilized by filtration.
Formulation
This PKLR protein is provided in a solution containing 20 mM Tris-HCl buffer (pH 8.0), 1 mM DTT, 0.2 M NaCl, and 10% glycerol. The protein concentration is 1 mg/ml.
Stability
For short-term storage (2-4 weeks), the protein can be stored at 4°C. For long-term storage, it is recommended to freeze the protein at -20°C. To prevent protein degradation during freezing and thawing, adding a carrier protein like HSA or BSA (0.1%) is advised. Repeated freezing and thawing should be avoided.
Purity
The purity of this PKLR protein is greater than 90%, as determined by SDS-PAGE analysis.
Biological Activity
The specific activity of this PKLR protein is >0.1 unit/mg. One unit of activity is defined as the amount of enzyme required to convert 1.0 µmol of phosphoenolpyruvate to pyruvate per minute at pH 7.5 and 37°C.
Synonyms
PK1, PKL, RPK, pyruvate kinase isozyme R/L, Red cell/liver pyruvate kinase, PKRL
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MLTQELGTAF FQQQQLPAAM ADTFLEHLCL LDIDSEPVAA RSTSIIATIG PASRSVERLK EMIKAGMNIA RLNFSHGSHE YHAESIANVR EAVESFAGSP LSYRPVAIAL DTKGPEIRTG ILQGGPESEV ELVKGSQVLV TVDPAFRTRG NANTVWVDYP NIVRVVPVGG RIYIDDGLIS LVVQKIGPEG LVTQVENGGV LGSRKGVNLP GAQVDLPGLS EQDVRDLRFG VEHGVDIVFA SFVRKASDVA AVRAALGPEG HGIKIISKIE NHEGVKRFDE ILEVSDGIMV ARGDLGIEIP AEKVFLAQKM MIGRCNLAGK PVVCATQMLE SMITKPRPTR AETSDVANAV LDGADCIMLS GETAKGNFPV EAVKMQHAIA REAEAAVYHR QLFEELRRAA PLSRDPTEVT AIGAVEAAFK CCAAAIIVLT TTGRSAQLLS RYRPRAAVIA VTRSAQAARQ VHLCRGVFPL LYREPPEAIW ADDVDRRVQF GIESGKLRGF LRVGDLVIVV TGWRPGSGYT NIMRVLSIS.
