L-Asparaginase 2

Description

Description
MW= 140 kDa calculated (tetramer). L-Asparaginase II from E. coli (ANSII) consists of a homotetrameric assembly of 140kDa with D2 symmetry. (residues 7-348, UniProtKB accession P00805).

Sequence  

        10         20         30         40         50
      TALA ALVMGFSGAA LALPNITILA TGGTIAGGGD SATKSNYTVG 
        60         70         80         90        100
KVGVENLVNA VPQLKDIANV KGEQVVNIGS QDMNDNVWLT LAKKINTDCD 
       110        120        130        140        150
KTDGFVITHG TDTMEETAYF LDLTVKCDKP VVMVGAMRPS TSMSADGPFN 
       160        170        180        190        200
LYNAVVTAAD KASANRGVLV VMNDTVLDGR DVTKTNTTDV ATFKSVNYGP 
       210        220        230        240        250
LGYIHNGKID YQRTPARKHT SDTPFDVSKL NELPKVGIVY NYANASDLPA 
       260        270        280        290        300
KALVDAGYDG IVSAGVGNGN LYKSVFDTLA TAAKTGTAVV RSSRVPTGAT 
       310        320        330        340 
TQDAEVDDAK YGFVASGTLN PQKARVLLQL ALTQTKDPQQ IQQIFNQY 

Purity
> 95% by SDS-PAGE. The protein is observed, in denaturing conditions, as a single band migrating at molecular weight at about 35.0 kDa.

Supplied as
1 mg/mL solution in phosphate buffer 150 mM pH 7.5. The concentration is calculated by the analysis of the absorbance at 280 nm (ε₂₈₀ = 23505 M^-1cm^-1 calculated for the monomer).
(unless otherwise specified, NMR tubes will be sold at a concentration of 1mM)

Characteristics
Under the above described conditions, the product can be concentrated up to 3 mM.

Storage
-20°C. The protein is stable at 4°C for months. Avoid repeated freeze/thaw cycles.
NMR USE: after the experiments return the sample at 4°C.

References
E. Ravera, S. Ciambellotti, L. Cerofolini, T. Martelli, T. Kozyreva, C. Bernacchioni, S. Giuntini, M. Fragai, P. Turano, C. Luchinat, Angew. Chem. Int. Ed. Engl. 2016,
55, 2446–2449.
L. Cerofolini, S. Giuntini, A. Carlon, E. Ravera, V. Calderone, M. Fragai, G. Parigi, C. Luchinat, Chemistry 2019, 25, 1984–1991.
S. Giuntini, L. Cerofolini, E. Ravera, M. Fragai, C. Luchinat, Sci Rep 2017, 7, 17934
S. Giuntini, E. Balducci, L. Cerofolini, E. Ravera, M. Fragai, F. Berti, C. Luchinat, Angew. Chem. Int. Ed. 2017, 56, 14997–15001.
L. Cerofolini, S. Giuntini, E. Ravera, C. Luchinat, F. Berti, M. Fragai, npj Vaccines 2019, 4, 20