The nuclear lamins form a karyoskeleton providing structural rigidity to the nucleus. One member of the lamin family, lamin A, is first synthesized as a 74 kDa precursor, prelamin A. After the endopeptidase and methylation reactions which occur after farnesylation of the CAAX-box cysteine, there is a second endoproteolysis that occurs 15 amino acids upstream from the C-terminal farnesylated cysteine residue. Studies with knockout mice have implicated the enzyme Zmpste24 (Face-1) as a suitable candidate to perform one or both of these proteolytic reactions. Evidence has been presented elsewhere establishing that Zmpste24 possesses a zinc-dependent CAAX endopeptidase activity. In the present study, we confirm this CAAX endopeptidase activity with recombinant, membrane-reconstituted Zmpste24 and show that it can accept a prelamin A farnesylated tetrapeptide as substrate. To monitor the second upstream endoproteolytic cleavage of prelamin A, we expressed a 33 kDa prelamin A C-terminal tail in insect cells. We demonstrate that this purified substrate possesses a C-terminal farnesylated and carboxyl-methylated cysteine and, therefore, constitutes a valid substrate for assaying the second endoproteolytic step in lamin A maturation. With this substrate, we demonstrate that insect cell membranes bearing recombinant Zmpste24 can also catalyse the second upstream endoproteolytic cleavage.

We perform sidebranch measurements on pure succinonitrile dendrites grown in both microgravity and terrestrial-gravity conditions for a set of supercoolings within the range 0.1-1.0 K. Two distinct types of sidebranch regions, uniform and coarsening, exist, and are characterized by the distance from the tip at which the region began, D(i), and the average spacing of sidebranches within that region, lambda(i). There does not appear to be any significant dependence on either gravity level or supercooling when D(i) or lambda(i) are normalized with respect to the radius of curvature of the tip, R. The apparently constant normalized proportionality factor between D(i), lambda(i), and R, regardless of the relative importance of diffusion and convective heat transport, demonstrates self-similarity between dendrites of different length scales propagating under various heat transfer conditions. However, when the form of the sidebranch envelope is defined by a power law relating the amplitude and relative positions of the sidebranches normalized to the radius of the tip, the form is seen to have significant variations with supercooling between the terrestrial gravity and microgravity growth dendrites. Furthermore, both the amplitude coefficient and exponent from the power-law regressions of the microgravity data are statistically different (95% confidence level) than their terrestrial counterparts.