PMID- 33533060
OWN - NLM
STAT- MEDLINE
DCOM- 20210325
LR  - 20210402
IS  - 1522-2683 (Electronic)
IS  - 0173-0835 (Print)
IS  - 0173-0835 (Linking)
VI  - 42
IP  - 6
DP  - 2021 Mar
TI  - Isoelectric point determination by imaged CIEF of commercially available 
      SARS-CoV-2 proteins and the hACE2 receptor.
PG  - 687-692
LID - 10.1002/elps.202100015 [doi]
AB  - In order to contribute to the scientific research on the severe acute respiratory 
      syndrome coronavirus 2 (SARS-CoV-2), we have investigated the isoelectric points 
      (pI) of several related proteins, which are commercially available: the 
      receptor-binding domain (RBD) with His- and Fc-tag, the S1 subunit with His-tag, 
      the S1/S2 subunits with His-tag and the human angiotensin-converting enzyme 2 
      (hACE2) with His-tag. First, the theoretical pI values, based on the amino acid 
      (AA) sequences of the proteins, were calculated using the ProtParam tool from the 
      Bioinformatics Resource Portal ExPASy. The proteins were then measured with the 
      Maurice imaged CIEF system (native fluorescence detection), testing various 
      measurement conditions, such as different ampholytes or ampholyte mixtures. Due 
      to isoforms, we get sections with several peaks and not just one peak for each 
      protein. The determined pI range for the RBD/Fc is 8.24-9.32 (theoretical pI: 
      8.55), for the RBD/His it is 7.36-9.88 (8.91) and for the S1/His it is 7.30-8.37 
      (7.80). The pI range of the S1/S2/His is 4.41-5.87 (no theoretical pI, AA 
      sequence unknown) and for hACE2/His, the determined global range is 5.19-6.11 
      (5.60) for all experimental conditions chosen. All theoretically derived values 
      were found within these ranges, usually close to the center. Therefore, we 
      consider theoretical values as useful to make predictions about the isoelectric 
      points of SARS-CoV-2 proteins. The experimental conditions had only a minor 
      influence on the pI ranges obtained and mainly influenced the peak shapes.
CI  - (c) 2021 The Authors. Electrophoresis published by Wiley-VCH GmbH.
FAU - Krebs, Finja
AU  - Krebs F
AD  - Institute of Medicinal and Pharmaceutical Chemistry, Technische Universitat 
      Braunschweig, Braunschweig, Germany.
FAU - Scheller, Christin
AU  - Scheller C
AD  - Institute of Medicinal and Pharmaceutical Chemistry, Technische Universitat 
      Braunschweig, Braunschweig, Germany.
FAU - Grove-Heike, Kristina
AU  - Grove-Heike K
AD  - Institute of Medicinal and Pharmaceutical Chemistry, Technische Universitat 
      Braunschweig, Braunschweig, Germany.
FAU - Pohl, Lena
AU  - Pohl L
AD  - Institute of Medicinal and Pharmaceutical Chemistry, Technische Universitat 
      Braunschweig, Braunschweig, Germany.
FAU - Watzig, Hermann
AU  - Watzig H
AUID- ORCID: 0000-0002-7456-0612
AD  - Institute of Medicinal and Pharmaceutical Chemistry, Technische Universitat 
      Braunschweig, Braunschweig, Germany.
LA  - eng
PT  - Journal Article
DEP - 20210212
PL  - Germany
TA  - Electrophoresis
JT  - Electrophoresis
JID - 8204476
RN  - 0 (Protein Subunits)
RN  - 0 (Spike Glycoprotein, Coronavirus)
RN  - 0 (spike protein, SARS-CoV-2)
RN  - EC 3.4.17.23 (ACE2 protein, human)
RN  - EC 3.4.17.23 (Angiotensin-Converting Enzyme 2)
SB  - IM
MH  - Angiotensin-Converting Enzyme 2/*chemistry/metabolism
MH  - Binding Sites
MH  - COVID-19/metabolism/*virology
MH  - Humans
MH  - Isoelectric Focusing/*methods
MH  - Isoelectric Point
MH  - Protein Interaction Domains and Motifs
MH  - Protein Subunits/chemistry/metabolism
MH  - SARS-CoV-2/*chemistry/metabolism
MH  - Spike Glycoprotein, Coronavirus/*chemistry/metabolism
PMC - PMC8013610
OTO - NOTNLM
OT  - CIEF
OT  - COVID-19
OT  - Isoelectric point
OT  - SARS-CoV-2 proteins
OT  - hACE2
COIS- The authors have declared no conflict of interest.
EDAT- 2021/02/04 06:00
MHDA- 2021/03/26 06:00
PMCR- 2021/04/01
CRDT- 2021/02/03 06:04
PHST- 2021/01/15 00:00 [received]
PHST- 2021/01/29 00:00 [revised]
PHST- 2021/02/01 00:00 [accepted]
PHST- 2021/02/04 06:00 [pubmed]
PHST- 2021/03/26 06:00 [medline]
PHST- 2021/02/03 06:04 [entrez]
PHST- 2021/04/01 00:00 [pmc-release]
AID - ELPS7365 [pii]
AID - 10.1002/elps.202100015 [doi]
PST - ppublish
SO  - Electrophoresis. 2021 Mar;42(6):687-692. doi: 10.1002/elps.202100015. Epub 2021 
      Feb 12.