Identification of a labelled peptide after stoicheiometric reaction of fluorescein isothiocyanate with the Ca2+‐dependent adenosine triphosphatase of sarcoplasmic reticulum

FEBS Letters

Mitchinson, Colin¹  Wilderspin, Andrew F.¹  Trinnaman, Brian J.¹  Green, N.Michael¹ 

[1] National Institute for Medical Research, Mill Hill, London NW7 1AA, England

关键词: (Ca2+ + Mg2+)ATPase inhibition;    Reactive lysine residue;    Nucleotide-binding fold;    Fluorescein isothiocyanate;    Integral membrane protein;    (Sarcoplasmic reticulum);    bicine;    N;    N-bis-(2-hydroxyethyl)-glycine;    Ca2+-ATPase;    calcium- and magnesium-dependent adenosine triphosphatase (EC 3.6.1.3);    C12E9;    nona-ethyleneglycol dodecyl ether;    FITC;    Fluorescein 5 isothiocyanate (isomer I);    FTC;    fluorescein thiocarbamyl;    SR;    sarcoplasmic reticulum;   

DOI  :  10.1016/0014-5793(82)80710-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Incorporation of 4.5 nmol fluorescein isothiocyanate/mg rabbit sarcoplasmic reticulum, or of 7.4 nmol/mg purified ATPase, was sufficient to inhibit the activity completely. These results are not consistent with the suggestion (Pick, U. and Karlish, S.J.D. (1980) Biochim. Biophys. Acta 626, 255–261) that 2 mol ATPase were inhibited by each mole of reagent incorporated. A single labelled peptide was purified from the inhibited ATPase and it was shown that Lys 3/190, 10 residues from the N-terminus of tryptic fragment B, was the reactive lysine residue. This site is close to a potential nucleotide-binding fold in the ATPase sequence. A similar peptide showing only 2 conservative replacements was isolated from the sarcoplasmic reticulum of the lobster.

【 授权许可】

Unknown