PMID- 35608746
OWN - NLM
STAT- MEDLINE
DCOM- 20220725
LR  - 20220725
IS  - 1573-4978 (Electronic)
IS  - 0301-4851 (Linking)
VI  - 49
IP  - 8
DP  - 2022 Aug
TI  - Bacterial over-expression of functionally active human CT2 (SLC22A16) carnitine 
      transporter.
PG  - 8185-8193
LID - 10.1007/s11033-022-07491-1 [doi]
AB  - BACKGROUND: Escherichia coli is a widely used tool for the over-expression of 
      human proteins for studying structure and function. The toxicity of human 
      proteins for E. coli often hampers the expression. This study aims to find 
      conditions for the expression of a membrane transporter known as the carnitine 
      transporter CT2. The knowledge on this transporter is scarce, thus obtaining the 
      recombinant protein is very important for further studies. METHODS AND RESULTS: 
      The cDNAs coding for human CT2 (hCT2) was cloned in the pH6EX3 vector and 
      different transformed E. coli strains were cultured in absence or in presence of 
      glucose. hCT2 expression was obtained. The protein was purified and reconstituted 
      into proteoliposomes in a functionally active state. CONCLUSIONS: Using the 
      appropriate IPTG concentration, together with the addition of glucose, hCT2 has 
      been expressed in E. coli. The protein is active and shows capacity to transport 
      carnitine in proteoliposomes. The results have a great interest in basic 
      biochemistry of membrane transporters and applications to human health since hCT2 
      is involved in human pathology.
CI  - (c) 2022. The Author(s), under exclusive licence to Springer Nature B.V.
FAU - Galluccio, Michele
AU  - Galluccio M
AD  - Department DiBEST Biologia, Ecologia, Scienze Della Terra Unit of Biochemistry 
      and Molecular Biotechnology, University of Calabria, Via P. Bucci cubo 4C, 87036, 
      Arcavacata di Rende, Italy.
FAU - Mazza, Tiziano
AU  - Mazza T
AD  - Department DiBEST Biologia, Ecologia, Scienze Della Terra Unit of Biochemistry 
      and Molecular Biotechnology, University of Calabria, Via P. Bucci cubo 4C, 87036, 
      Arcavacata di Rende, Italy.
FAU - Scalise, Mariafrancesca
AU  - Scalise M
AD  - Department DiBEST Biologia, Ecologia, Scienze Della Terra Unit of Biochemistry 
      and Molecular Biotechnology, University of Calabria, Via P. Bucci cubo 4C, 87036, 
      Arcavacata di Rende, Italy.
FAU - Sarubbi, Maria Chiara
AU  - Sarubbi MC
AD  - Department DiBEST Biologia, Ecologia, Scienze Della Terra Unit of Biochemistry 
      and Molecular Biotechnology, University of Calabria, Via P. Bucci cubo 4C, 87036, 
      Arcavacata di Rende, Italy.
FAU - Indiveri, Cesare
AU  - Indiveri C
AUID- ORCID: 0000-0001-9818-6621
AD  - Department DiBEST Biologia, Ecologia, Scienze Della Terra Unit of Biochemistry 
      and Molecular Biotechnology, University of Calabria, Via P. Bucci cubo 4C, 87036, 
      Arcavacata di Rende, Italy. cesare.indiveri@unical.it.
AD  - CNR Institute of Biomembranes, Bioenergetics and Molecular Biotechnology IBIOM, 
      via Amendola 122/O, 70126, Bari, Italy. cesare.indiveri@unical.it.
LA  - eng
GR  - PON ARS01_00568/Ministero dell'Istruzione, dell'Universita e della Ricerca/
PT  - Journal Article
DEP - 20220524
PL  - Netherlands
TA  - Mol Biol Rep
JT  - Molecular biology reports
JID - 0403234
RN  - 0 (Membrane Transport Proteins)
RN  - 0 (Organic Cation Transport Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (SLC22A16 protein, human)
RN  - IY9XDZ35W2 (Glucose)
RN  - S7UI8SM58A (Carnitine)
SB  - IM
MH  - *Carnitine/metabolism
MH  - *Escherichia coli/genetics/metabolism
MH  - Glucose/metabolism
MH  - Humans
MH  - Membrane Transport Proteins/metabolism
MH  - Organic Cation Transport Proteins/genetics
MH  - Recombinant Proteins/metabolism
OTO - NOTNLM
OT  - CT2
OT  - Carnitine
OT  - Liposome
OT  - Protein expression
OT  - Purification
OT  - SLC
EDAT- 2022/05/25 06:00
MHDA- 2022/07/26 06:00
CRDT- 2022/05/24 11:19
PHST- 2021/11/11 00:00 [received]
PHST- 2022/04/20 00:00 [accepted]
PHST- 2022/05/25 06:00 [pubmed]
PHST- 2022/07/26 06:00 [medline]
PHST- 2022/05/24 11:19 [entrez]
AID - 10.1007/s11033-022-07491-1 [pii]
AID - 10.1007/s11033-022-07491-1 [doi]
PST - ppublish
SO  - Mol Biol Rep. 2022 Aug;49(8):8185-8193. doi: 10.1007/s11033-022-07491-1. Epub 
      2022 May 24.