Table 1.

Summary of the assembly properties of K14WT and the K14CF and K14CF-C367 variants

Settings Features K14 variant analyzed 
  K14WT K14CF K14CF-C367 
In vitroa (purified proteins)     
Reducing conditions IF length Long Long Long 
 IF width Normal Normal Normal 
 Assembly efficiency High High High 
Oxidizing conditions IF length Very short Intermediate Intermediate 
 IF width Normal Normal Aberrant 
 Assembly efficiency Reduced High Reduced 
Oxidizing→reducing IF length Long Long Long 
 IF width Normal Normal Normal 
 Assembly efficiency High High High 
Krt14−/− newborn skin keratinocytes in primary cultureb Frequency of perinuclear-concentrated networks High Low Significantly restored (i.e. wild type–like) 
 Frequency of pan-cytoplasmic networks Low High Significantly restored 
 Keratin cycling Normal Impaired 45%: Significantly restored55%: Impaired 
 Movement of nucleus and whole cell Limited Exaggerated Significantly restored 
Settings Features K14 variant analyzed 
  K14WT K14CF K14CF-C367 
In vitroa (purified proteins)     
Reducing conditions IF length Long Long Long 
 IF width Normal Normal Normal 
 Assembly efficiency High High High 
Oxidizing conditions IF length Very short Intermediate Intermediate 
 IF width Normal Normal Aberrant 
 Assembly efficiency Reduced High Reduced 
Oxidizing→reducing IF length Long Long Long 
 IF width Normal Normal Normal 
 Assembly efficiency High High High 
Krt14−/− newborn skin keratinocytes in primary cultureb Frequency of perinuclear-concentrated networks High Low Significantly restored (i.e. wild type–like) 
 Frequency of pan-cytoplasmic networks Low High Significantly restored 
 Keratin cycling Normal Impaired 45%: Significantly restored55%: Impaired 
 Movement of nucleus and whole cell Limited Exaggerated Significantly restored 
a

(1) Formation of disulfide bonds limits the elongation of filaments; this effect is fully reversible. (2) When present as the only cysteine residue in K14, C367 interferes with elongation and causes a dramatic loss of filament width control.

b

(1) K14-bound cysteines and/or disulfide bonds are required for the centripetal keratin cycle to occur in live cells. (2) When present as the only cysteine residue in K14, C367 significantly rescues the keratin cycle, even though it exhibits aberrant properties in vitro.

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