Table 7.
Expression of MCCC1 and MCCC2 wildtype and mutant alleles
| Allele |
PCC and MCC activities (pmol/min/mg protein)* |
|||||
|---|---|---|---|---|---|---|
|
Experiment 1 |
Experiment 2 |
|||||
| PCC | MCC | %** | PCC | MCC | %** | |
| MCCC1-wildtype |
311 |
193 |
100 |
331 |
157 |
100 |
| vector only |
335 |
0 |
0 |
377 |
0.3 |
0.2 |
| MCCC1-p.E288G |
283 |
0 |
0 |
352 |
1.8 |
1.1 |
| MCCC1-p.G379D |
293 |
0 |
0 |
326 |
0 |
0 |
| MCCC1-p.I434M |
330 |
87.6 |
45.4 |
324 |
74.5 |
47.4 |
| | ||||||
| MCCC2-wildtype |
377 |
75.4 |
100 |
341 |
49.4 |
100 |
| vector only |
364 |
0.3 |
0.4 |
346 |
0 |
0 |
| MCCC2-p.S39F |
398 |
29.6 |
39.3 |
368 |
24.8 |
50.2 |
| MCCC2-p.G118del |
366 |
10.8 |
14.3 |
342 |
3.2 |
6.5 |
| MCCC2-p.Y146N |
341 |
58.7 |
77.9 |
372 |
44.1 |
89.3 |
| MCCC2-p.H282R |
301 |
9.6 |
12.7 |
339 |
2.3 |
4.7 |
| MCCC2-p.A456V |
265 |
2.0 |
2.7 |
340 |
0.1 |
0.2 |
| MCCC2-p.S523G |
313 |
56.1 |
74.4 |
344 |
30.9 |
62.6 |
| | ||||||
| MCCC2-wildtype |
335 |
76.9 |
100 |
446 |
43.5 |
100 |
| vector only |
371 |
0 |
0 |
540 |
0.5 |
1.1 |
| MCCC2-p.V434L |
295 |
56.6 |
73.7 |
449 |
33.5 |
77.0 |
| MCCC2-p.G475R | 290 | 33.7 | 43.8 | 414 | 21.1 | 48.5 |
Transient expression was performed in transformed MCCC1 and MCCC2 deficient fibroblasts followed by the assay of propionyl-CoA carboxylase (PCC) and 3-methylcrotonyl-CoA carboxylase (MCC) activities. Transfection with an empty vector (vector only) was used as negative control.
*activities are the mean of duplicate determination.
** % of MCC activity of simultaneously expressed wildtype allele.