The SDS-PAGE containing connexin 43 is shown in Figure 5. There was a twofold increase in radioactivity compared with the supernatant, which was not different from background activity. In the next step of the further purification of the connexins, ie, of con-nexin 43, radioactivity was lost. Finally, connexin 43 was found, but no 14C-labelled AAP10 was detectable (Table 2) Antiarrhythmic peptides: A new antiarrhythmic principle

Figure 5 SDS-Page of purified gap junctional membranes. Numbers indicate pufication steps. GB Gradient band, ie, the interface material of step 36; GP Gradient pellet; M Molecular weight marker. Lanes to the left of M were derivedfrom undiluted test volumes; lanes to the right of M were diluted 1:1 with buffer solution. Note the 43 kDa band containing connexin 43 from rabbit hearts. During further purification, connexin 43 was enriched in the gradient band
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TABLE 2 Radioactivity in pellets and supernatants following purification of connexin 43

Fraction Radioactivity(counts/min) Comment
Standard 107,300.00
Perfusate 58,977.00
Supernatant S9 11,962.03
Pellet S9 632.83
Supernatant S11 162.97
Pellet S11 153.61
Supernatant S16 62.12
Pellet S16 89.78
Supernatant S21 39.82
Pellet S21 83.30 Intracellular and membrane-associated proteins
H2O 38.85
Connexin 43 nd Radiofluorography/PAGEgel electrophoresis

nd None detectable. Numbers (Sx) refer to the step numbers given in the Methods section. Note that there was increased activity in the pellet of step 21

Category: Cardiology / Tags: Antiarrhythmic drugs, Antiarrhythmic peptides, Cardiac arrhythmia, Cellular coupling, Gap junction

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