Precise comparisons of mammalian gene maps require common anchor loci as landmarks for conserved chromosomal segments. Using a computer script that automates DNA sequence database alignments, we designed 410 evolutionarily conserved primer pair sequences which are specific for anchor locus gene amplification from any mammalian species' DNA. Primer pairs were designed to span introns for polymorphism ascertainment, and to include sufficient exonic sequence (25-400 bp) to allow for gene identification. A total of 318 primer pairs were optimized for domestic cats, and 86% of the sequenced feline PCR products showed homology to the gene of primer origin. A screen of 20 mammals from 11 orders revealed that 35-52% of the 318 primers yielded a single PCR product without further optimization suggesting that nearly 75% can be optimized for any eutherian mammal.

The measurement of clonal heterogeneity is central to understanding the evolutionary and population genetics of the roughly 50 species of vertebrates that lack effective genetic recombination.

Previously melittin, the α-helical basic honey bee venom peptide, was shown to inhibit F 1 -ATPase by binding at the β-subunit DELSEED motif of F 1 F o ATP synthase. Herein, we present the inhibitory effects of the basic α-helical amphibian antimicrobial peptides, ascaphin-8, aurein 2.2, aurein 2.3, carein 1.8, carein 1.9, citropin 1.1, dermaseptin, maculatin 1.1, maganin II, MRP, or XT-7, on purified F 1 and membrane bound F 1 F o E. coli ATP synthase. We found that the extent of inhibition by amphibian peptides is variable. Whereas MRP-amide inhibited ATPase essentially completely (~96% inhibition), carein 1.8 did not inhibit at all (0% inhibition). Inhibition by other peptides was partial with a range of ~13% to 70%. MRP-amide was also the most potent inhibitor on molar scale (IC 50 ~3.25 µM). Presence of an amide group at the c-terminal of peptides was found to be critical in exerting potent inhibition of ATP synthase (~20-40% additional inhibition). Inhibition was fully reversible and found to be identical in both F 1 F o membrane preparations as well as in isolated purified F 1 . Interestingly, growth of Escherichia coli was abrogated in the presence of ascaphin-8, aurein 2.2, aurein 2.3, citropin 1.1, dermaseptin, magainin II-amide, MRP, MRP-amide, melittin, or melittinamide but was unaffected in the presence of carein 1.8, carein 1.9, maculatin 1.1, magainin II, or XT-7. Hence inhibition of F 1 -ATPase and E. coli cell growth by amphibian antimicrobial peptides suggests that their antimicrobial/anticancer properties are in part linked to their actions on ATP synthase. KeywordsF 1 F o -ATP synthase; F 1 -ATPase; E. coli ATP synthase; Antimicrobial peptides; Amphibian; Enzyme inhibitors F 1 F o -ATP synthase is the primary source of cellular energy production in animals, plants, and almost all microorganisms by oxidative or photophosphorylation. The ATP synthase enzyme is highly conserved and structurally similar in all organisms. This enzyme is the smallest known biological nanomotor and is composed of two rotary sectors, F 1 and F o . In its simplest form in Escherichia coli ATP synthase contains eight different subunits namely α 3 ß 3 γδεab 2 c 10 with a total molecular mass of ~530 kDa. F 1 corresponds to α 3 ß 3 γδε -and F o to ab 2 c 10 . The reversible processess of ATP hydrolysis and synthesis occur on three catalytic sites in the F 1 sector, Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. whereas proton transport occurs through the membrane embedded F o [1][2]. An important feature of the molecular mechanism of ATP synthase is that a "rotor" made up...

Recombination is unknown in natural popu-

In this review we discuss the inhibitory effects of dietary polyphenols and amphibian antimicrobial/antitumor peptides on ATP synthase. In the beginning general structural features highlighting catalytic and motor functions of ATP synthase will be described. Some details on the presence of ATP synthase on the surface of several animal cell types, where it is associated with multiple cellular processes making it an interesting drug target with respect to dietary polyphenols and amphibian antimicrobial peptides will also be reviewed. ATP synthase is known to have distinct polyphenol and peptide binding sites at the interface of α/β subunits. Molecular interaction of polyphenols and peptides with ATP synthase at their respective binding sites will be discussed. Binding and inhibition of other proteins or enzymes will also be covered so as to understand the therapeutic roles of both types of molecules. Lastly, the effects of polyphenols and peptides on the inhibition of Escherichia coli cell growth through their action on ATP synthase will also be presented.