CHAPTER 24

Lead-Calcium Interactions and Lead Toxicity

T. J. B. SIMONS

A. Overview
The aim of this chapter is to bring together information on the interactions be-
tween Ca and Pb at the cellular level, and to discuss them in relation to possible
mechanisms for the toxicity of Pb. Interactions between Pb and Ca in the whole
organism have been known for many years. For example, Ca inhibits Pb uptake
in the gastrointestinal tract, and long-term storage of Pb occurs in the bones (So-
BEL et al. 1940; SMITH and HURSH 1977; MAHAFFEY 1980). These processes are
now being studied at the cellular and molecular level. Interactions with binding
proteins, enzymes, membrane transport, and secretory mechanisms will be discus-
sed in later sections of this chapter. Consideration of the organism as a whole will
be excluded, as will the interaction of Pb with other metals. (For interactions with
Fe, Cu and Zn, see, for example MAHAFFEY and RADER 1980; PETERING 1980.)
The traditional view of the toxicology of Pb and other heavy metals is that
they inhibit enzymes by interaction with SH groups (VALLEE and ULMER 1972).
An alternative hypothesis has recently been suggested, based upon the observa-
tion that Pb and other heavy metals interact with calmodulin. Calmodulin is
found in all cells and acts as a Ca 2 + receptor, mediating many of the intracellular
effects of Ca 2 + ions. Lead can replace Ca 2 + and causes activation of calmodulin
at low concentrations, and inhibition at high concentrations (CHAO et al. 1984).
It has been suggested that the toxic effects of Pb and other heavy metals might
be brought about by: (a) occupying Ca-binding sites on calmodulin (GOLDSTEIN
and AR 1983); (b) inhibiting calmodulin (Cox and HARRISON 1983); or (c) activat-
ing calmodulin (CHEUNG 1984). It will be shown that this view is oversimplified,
because there are other Pb-Ca interactions at the cellular and molecular level, and
because Pb has other actions that are unlike those of Ca. These include enzyme
inhibition, binding to specific proteins, and a novel mechanism of membrane
transport. The conclusion will be that Pb toxicity is due to a combination of ef-
fects, some like and some unlike those of Ca.

B. Relevant Chemistry of Lead and Calcium

I. Introduction
Lead, atomic number 82, is in group IVB of the periodic table. It exists in two
oxidation states, Pb(II) and Pb(IV), but only Pb(II) is normally encountered in
nature. Inorganic Pb(IV) compounds are unstable under physiological condi-
P. F. Baker (ed.), Calcium in Drug Actions
© Springer-Verlag Berlin Heidelberg 1988
510 T. J. B. SIMONS

tions. One organic Pb(IV) compound, tetraethyllead, is widely used as a petro-

leum additive, but it is converted to Pb(II) during or after combustion. Pb occurs
naturally as the minerals PbS and PbC0 3 . Lead oxide (PbO) forms salts with
many acids. These salts contain the Pb 2 + ion, which is the form in which Pb
usually interacts with biological systems.
Calcium, atomic number 20, is an alkaline earth (group IIA). It only exists in
one oxidation state, Ca(U), and is widely distributed in nature as CaC0 3 and
CaS0 4 . It is an essential constituent ofliving matter, where it is always found as
the Ca 2 + ion, either in solution, or in complexed or mineral form.

II. Chemistry of the Ions in Solution

1. Complexes with Simple Anions
The Pb 2 + ions has two 6s electrons in its outer valence shell, and also low-lying
unfilled p and d orbitals. It can use these to form covalent or partial ionic/covalent
bonds with other ions. Complexes are formed with many inorganic anions in so-
lution, e.g. with OH-, CI- and NO;. Successive addition of an anion (X-) leads
to formation of PbX+, PbX 2 , PbX; and PbXi - species (SILLEN and MARTELL
1964). Other, more complex ions can also be formed. The neutral species often
has a low water solubility, and this can limit the range of Pb2+ concentrations
that can be achieved experimentally. Addition of Pb in increasing quantities to
physiological solutions that mimic mammalian extracellular fluid leads to precip-
itation in the sequence Pb 3 (P0 4 b PbC0 3 then Pb(OH)z (MAXWELL and
BISCHOFF 1929; SIMONS 1986a). The solubility product of Pb(OH)z (1.7 x 10- 19
M3; BIRRAUX et al. 1977) imposes an upper limit of about (5-10) x 10- 6 Mas
the maximum Pb 2 + concentration that can be achieved in vitro at neutral pH.
Many publications report the use of much higher concentrations of lead nitrate
or acetate. These experiments should be interpreted with considerable caution.
All the valence shells on Ca 2 + are full, and it does not form significant complexes
with inorganic anions under normal conditions.

2. Complexes with Organic Ligands

Ca2+ and Pb2+ both form complexes with chelating ligands, such as ethylenedi-
amine tetraacetic acid and nitrilotriacetic acid. The binding of Pb 2 + is always
stronger than that of Ca 2 +, because of its orbital structure. The difference in af-
finity depends upon the donor atoms present at the binding site. Pb 2 + has a much
greater tendency to form bonds to Nand S ligands than Ca2+, and the presence
of these ligands leads to a greatly increased binding of Pb 2 + compared with
Ca 2 +.
Ca 2 + is always coordinated by oxygen atoms in proteins containing specific
Ca-binding sites. The coordination number is 6, 7 or 8, and the oxygens are in
carbonyl or carboxyl groups, or water molecules (KRETSINGER 1976; MARTIN
1984). It seems reasonable to assume that Pb 2 + binds at these sites when it pro-
duces Ca-like effects. In view of its tendency to form bonds to Nand S, it prob-
ably also binds to other sites in proteins, where these may be present. The use of
Pb derivatives in the determination of crystal structures provides some informa-