We have designed a heterodimerizing leucine zipper system to target a radio
nuclide to prelocalized non-internalizing tumor-specific antibodies. The mo
dular nature of the leucine zipper allows us to iteratively use design rule
s to achieve specific homodimer and heterodimer affinities. We present circ
ular-dichroism thermal denaturation measurements on four pairs of heterodim
erizing leucine zippers. These peptides are 47 amino acids long and contain
four or five pairs of electrostatically attractive g <----> e ' (i, i ' +5
) interhelical heterodimeric interactions. The most stable heterodimer cons
ists of an acidic leucine zipper and a basic leucine zipper that melt as ho
modimers in the micro (T-m = 28 degreesC) or nanomolar (T-m = 40 degreesC)
range, respectively, but heterodimerize with a T-m >90 degreesC, calculated
to represent femtamolar affinities. Modifications to this pair of acidic a
nd basic zippers, designed to destabilize homodimerization, resulted in pep
tides that are unstructured monomers at 4 muM and 6 degreesC but that heter
odimerize with a T-m = 74 degreesC or K-d(37) = 1.1 x 10(-11) M. A third he
terodimerizing pair was designed to have a more neutral isoelectric focusin
g point (pI) and formed a heterodimer with T-m = 73 degreesC. We can tailor
this heterodimerizing system to achieve pharmacokinetics aimed at optimizi
ng targeted killing of cancer cells.