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Types of neurons, synaptic connections and chemical characteristics of cells immunoreactive for calbindin-D28K, parvalbumin and calretinin in the neocortex.

This article provides a general account of types of neurons, synaptic connections and chemical characteristics (colocalization studies) of cells immunoreactive for the three main calcium-binding proteins found in the neocortex, namely, calbindin-D28K, parvalbumin and calretinin. The main conclusion is two-fold. First, all, or the majority, of calbindin-, parvalbumin- and calretinin-immunoreactive cells are smooth nonpyramidal neurons (interneurons) which participate in a variety of complex cortical circuits that may differ depending on the species, cortical area or layer where they are located. Second, in general, different types of nonpyramidal neurons are stained for each of these calcium-binding proteins and display different chemical characteristics regarding a variety of neurotransmitters (or related compounds), cell surface markers and receptors. However, a certain overlap exits, which also shows regional and species differences.

Adult↗

Comparative analysis of calcium-binding protein-immunoreactive neuronal populations in the auditory and visual systems of the bottlenose dolphin (Tursiops truncatus) and the macaque monkey (Macaca fascicularis).

This study compares the distribution of three calcium-binding protein-immunoreactive (CaBP-immunoreactive) neuronal populations (calretinin-, calbindin- and parvalbumin-immunoreactive) in the visual and auditory systems in two mammalian species which are fundamentally different in their evolutionary traits and ecology, the aquatic toothed whale Tursiops truncatus (bottlenose dolphin) and the terrestrial Old World primate, Macaca fascicularis (long-tailed macaque). Immunocytochemical analyses, combined with computerized morphometry revealed that in the visual and auditory systems of the bottlenose dolphin, calretinin and calbindin are the prevalent calcium-binding proteins, whereas parvalbumin is present in very few neurons. The prevalence of calretinin and calbindin-immunoreactive neurons is especially obvious in the auditory system of this species. In both auditory and visual systems of the macaque monkey, the parvalbumin-immunoreactive neurons are present in comparable or higher densities than the calretinin and calbindin-immunoreactive neurons. In some structures of the visual and auditory systems of the macaque monkey, the calretinin- and calbindin-immunoreactive neurons are nearly absent. The prevalence of parvalbumin-immunoreactive over calretinin- and calbindin-immunoreactive neurons is particularly prominent in the visual system of primates. Thus, the dominant sensory systems in both aquatic and terrestrial mammals are enriched in specific phenotypes of calcium-binding protein-immunoreactive neurons.

Animals↗

Chemoarchitecture of the primate dorsal raphe nucleus.

The dorsal raphe nucleus (DR) harbours the largest single collection of serotonin (5-HT)-containing neurons in the brain but also comprises other types of chemospecific neurons. The aim of the present study was to characterise morphologically and immunohistochemically the DR in the squirrel monkey (Saimiri sciureus). The morphology of the DR 5-HT-immunoreactive (ir) neurons was analysed and their distribution compared to that of neurons displaying immunoreactivity for either tyrosine hydroxylase (TH), gamma-aminobutyric acid (GABA), substance P (SP), calbindin-D28k (CB), calretinin (CR) or parvalbumin (PV). The 5-HT-ir neurons were distributed in a highly heterogeneous manner throughout the rostrocaudal extent of the DR. The morphology and density of the 5-HT neurons were found to vary significantly in the major subdivisions of the primate DR, that is, the median, ventral, dorsal, ventrolateral, lateral and caudal subnuclei. Numerous SP-, GABA- and PV-ir neurons occurred in all six subnuclei of the DR. The distribution of SP-ir neurons was largely in register with that of 5-HT-ir neurons. Neurons expressing the other neuronal markers (TH, CB, CR) were not present in all six DR subnuclei and their distribution was either complementary to, or in register with, that of 5-HT-ir neurons. The median subnucleus was unique because it contained all the different types of chemospecific neurons. This study has revealed that the primate DR is chemically highly heterogeneous, a finding that may explain the multifarious influence that this nucleus exerts upon various forebrain structures.

Animals↗

Cellular distribution of the calcium-binding proteins parvalbumin, calbindin, and calretinin in the neocortex of mammals: phylogenetic and developmental patterns.

The three calcium-binding proteins parvalbumin, calbindin, and calretinin are found in morphologically distinct classes of inhibitory interneurons as well as in some pyramidal neurons in the mammalian neocortex. Although there is a wide variability in the qualitative and quantitative characteristics of the neocortical subpopulations of calcium-binding protein-immunoreactive neurons in mammals, most of the available data show that there is a fundamental similarity among the mammalian species investigated so far, in terms of the distribution of parvalbumin, calbindin, and calretinin across the depth of the neocortex. Thus, calbindin- and calretinin-immunoreactive neurons are predominant in layers II and III, but are present across all cortical layers, whereas parvalbumin-immunoreactive neurons are more prevalent in the middle and lower cortical layers. These different neuronal populations have well defined regional and laminar distribution, neurochemical characteristics and synaptic connections, and each of these cell types displays a particular developmental sequence. Most of the available data on the development, distribution and morphological characteristics of these calcium-binding proteins are from studies in common laboratory animals such as the rat, mouse, cat, macaque monkey, as well as from postmortem analyses in humans, but there are virtually no data on other species aside of a few incidental reports. In the context of the evolution of mammalian neocortex, the distribution and morphological characteristics of calcium-binding protein-immunoreactive neurons may help defining taxon-specific patterns that may be used as reliable phylogenetic traits. It would be interesting to extend such neurochemical analyses of neuronal subpopulations to other species to assess the degree to which neurochemical specialization of particular neuronal subtypes, as well as their regional and laminar distribution in the cerebral cortex, may represent sets of derived features in any given mammalian order. This could be particularly interesting in view of the consistent differences in neurochemical typology observed in considerably divergent orders such as cetaceans and certain families of insectivores and metatherians, as well as in monotremes. The present article provides an overview of calcium-binding protein distribution across a large number of representative mammalian species and a review of their developmental patterns in the species where data are available. This analysis demonstrates that while it is likely that the developmental patterns are quite consistent across species, at least based on the limited number of species for which ontogenetic data exist, the distribution and morphology of calcium-binding protein-containingneurons varies substantially among mammalian orders and that certain species show highly divergent patterns compared to closely related taxa. Interestingly, primates, carnivores, rodents and tree shrews appear closely related on the basis of the observed patterns, marsupials show some affinities with that group, whereas prototherians have unique patterns. Our findings also support the relationships of cetaceans and ungulates, and demonstrates possible affinities between carnivores and ungulates, as well as the existence of common, probably primitive, traits in cetaceans and insectivores.

Animals↗

FAK deficiency in cells contributing to the basal lamina results in cortical abnormalities resembling congenital muscular dystrophies.

Targeted deletion of focal adhesion kinase (fak) in the developing dorsal forebrain resulted in local disruptions of the cortical basement membrane located between the neuroepithelium and pia-meninges. At disruption sites, clusters of neurons invaded the marginal zone. Retraction of radial glial endfeet, midline fusion of brain hemispheres, and gliosis also occurred, similar to type II cobblestone lissencephaly as seen in congenital muscular dystrophy. Interestingly, targeted deletion of fak in neurons alone did not result in cortical ectopias, indicating that fak deletion from glia is required for neuronal mislocalization. Unexpectedly, fak deletion specifically from meningeal fibroblasts elicited similar cortical ectopias in vivo and altered laminin organization in vitro. These observations provide compelling evidence that FAK plays a key signaling role in cortical basement membrane assembly and/or remodeling. In addition, FAK is required within neurons during development because neuron-specific fak deletion alters dendritic morphology in the absence of lamination defects.

Animals↗

A novel calcium-sensitive switch revealed by the structure of human S100B in the calcium-bound form.

BACKGROUND: S100B is a homodimeric member of the EF-hand calcium-binding protein superfamily. The protein has been implicated in cellular processes such as cell differentiation and growth, plays a role in cytoskeletal structure and function, and may have a role in neuropathological diseases, such as Alzheimers. The effects of S100B are mediated via its interaction with target proteins. While several studies have suggested that this interaction is propagated through a calcium-induced conformational change, leading to the exposure of a hydrophobic region of S100B, the molecular details behind this structural alteration remain unclear. RESULTS: The solution structure of calcium-saturated human S100B (Ca(2+)-S100B) has been determined by heteronuclear NMR spectroscopy. Ca(2+)-S100B forms a well defined globular structure comprising four EF-hand calcium-binding sites and an extensive hydrophobic dimer interface. A comparison of Ca(2+)-S100B with apo S100B and Ca(2+)-calbindin D9k indicates that while calcium-binding to S100B results in little change in the site I EF-hand, it induces a backbone reorientation of the N terminus of the site II EF-hand. This reorientation leads to a dramatic change in the position of helix III relative to the other helices. CONCLUSIONS: The calcium-induced reorientation of calcium-binding site II results in the increased exposure of several hydrophobic residues in helix IV and the linker region. While following the general mechanism of calcium modulatory proteins, whereby a hydrophobic target site is exposed, the 'calcium switch' observed in S100B appears to be unique from that of other EF-hand proteins and may provide insights into target specificity among calcium modulatory proteins.

Amino Acids↗

Transient calretinin expression defines early postmitotic step of neuronal differentiation in adult hippocampal neurogenesis of mice.

We here show that the early postmitotic stage of granule cell development during adult hippocampal neurogenesis is characterized by the transient expression of calretinin (CR). CR expression was detected as early as 1 day after labeling dividing cells with bromodeoxyuridine (BrdU), but not before. Staining for Ki-67 confirmed that no CR-expressing cells were in cell cycle. Early after BrdU, CR colocalized with immature neuronal marker doublecortin; and later with persisting neuronal marker NeuN. BrdU/CR-labeled cells were negative for GABA and GABAA1 receptor, but early on expressed granule cell marker Prox-1. After 6 weeks, no new neurons expressed CR, but all contained calbindin. Stimuli inducing adult neurogenesis have limited (enriched environment), strong (voluntary wheel running), and very strong effects on cell proliferation (kainate-induced seizures). In these models the induction of cell proliferation was paralleled by an increase of CR-positive cells, indicating the stimulus-dependent progression from cell division to a postmitotic stage.

Aging↗

Blot analyses and immunocytochemistry of neural antigens with digoxigenylated primary and secondary antibodies.

While the digoxigenin-anti-digoxigenin (DIG) method is currently the preferred tool for non-radioactive in situ hybridization this study extends its application field to Western blotting of proteins and summarizes advantageous properties of digoxigenylated antibodies in immunocytochemistry. An established protocol for the preparation of digoxigenylated primary antibodies is complemented by dot blot analyses confirming the high sensitivity of hapten-anti-hapten techniques based on primary digoxigenylated antibodies. The comparative Western blot analysis of calcium-binding proteins in nervous tissue is used as an example to show the highly specific detection of relevant antigens with unmodified primary antibodies, digoxigenylated secondary antibodies and anti-digoxigenin-peroxidase conjugates. The application of the DIG technology seems to be especially indicated in tissues containing high amounts of endogenous biotin-bearing proteins which might induce false-positive staining in conventional streptavidin/biotin techniques. Finally, the previously shown suitability of digoxigenylated antibodies for different immunocytochemical procedures is completed here by examples for sensitive single immunoperoxidase staining of neural markers in rat brain and for carbocyanine double immunofluorescence labelling of senile plaques in old rhesus monkeys.

Animals↗

Chemical coding of sympathetic neurons controlling the tarsal muscle of the rat.

Sympathetic axons in the upper eyelid and in tissues in the superior retro-orbital space were examined for NPY immunoreactivity. Sympathetic nerve terminals containing co-localised NPY were associated with blood vessels, the conjunctiva and the Meibomian glands. The acini of the Harderian gland completely lacked sympathetic innervation. Sympathetic axons lacking NPY were only found in the tarsal muscle. In addition, a minority of terminals, located in the more proximal part of the tarsal muscle, contained weak immunoreactivity to NPY. Injections of the retrograde tracer, Fast Blue, into the eyelid or retro-orbital space labelled postganglionic somata in the superior cervical ganglion. While many retrogradely labelled somata were immunoreactive for NPY, around half lacked NPY immunoreactivity and so are likely to project to the tarsal muscle. Most of the retrogradely labelled postganglionic somata lacking NPY were surrounded by terminals immunoreactive for met-enkephalin, leu-enkephalin and met-enkephalin arg-gly-leu which were all found to be present in the same nerve terminals. Sectioning the cervico-sympathetic trunk eliminated all enkephalin-immunoreactive pericellular baskets. Many enkephalin-immunoreactive pericellular terminals contained co-localised VAChT, calretinin and calbindin immunoreactivity, but completely lacked nitric oxide synthase immunoreactivity. A second population of nerve terminals that were immunoreactive for nitric oxide synthase also surrounded tarsal muscle-projecting neurons, but these terminals lacked immunoreactivity to enkephalin. Thus, postganglionic neurons projecting to the tarsal muscle are of at least two chemical phenotypes (with or without NPY) and they receive convergent input from at least two populations of preganglionic neurons with distinctive chemical phenotypes.

Amidines↗

Calcium-binding proteins immunoreactivity in the human subcortical and cortical visual structures.

The distribution of neurons and fibers immunoreactive (ir) to the three calcium-binding proteins parvalbumin (PV), calbindin D-28k (CB), and calretinin (CR) was studied in the human lateral geniculate nucleus (LGN), lateral inferior pulvinar, and optic radiation, and related to that in the visual cortex. In the LGN, PV, CR, and CB immunoreactivity was present in all laminae, slightly stronger in the magnocellular than in the parvocellular laminae for CB and CR. PV-ir puncta, representing transversally cut axons, and CR-ir fibers were revealed within the laminae and interlaminar zones, and just beyond the outer border of lamina 6 in the geniculate capsule. In the optic radiation both PV- and CR-immunoreactive neurons, puncta, and fibers were present. CB immunoreactivity was revealed in neurons of all laminae of the lateral geniculate nucleus, including S lamina and interlaminar zones. There were hardly any CB-ir puncta or fibers in the laminae, interlaminar zones, geniculate capsule, or optic radiation. In the lateral inferior pulvinar, immunoreactive neurons for the three calcium-binding proteins were present in smaller number than in the LGN, as well as PV-ir puncta and CR-ir fibers within the nucleus and in the pulvinar capsule. In the white matter underlying area 17, fibers intermingled with a few scattered neurons were stained for both PV and CR, but very rarely for CB. These fibers stopped at the limit between areas 17 and 18. Area 17 showed a dense plexus of PV-ir puncta and neurons in the thalamo-receptive layer IV and CR-ir puncta and neurons both in the superficial layers I-II, IIIC, and in layer VA. Cajal-Retzius CR-ir neurons were present in layer I. CB-ir puncta were almost confined to layer I-III and CB-ir neurons to layer II. Finally the superior colliculus exhibited mostly populations of PV and CR pyramidal-like immunoreactive neurons, mainly in the intermediate tier. These data suggest that in the visual thalamus most calcium-binding protein immunoreactive neurons project to the visual cortex, while in the superior colliculus a smaller immunoreactive population represent projection neurons.

Adult↗

Calretinin in the cat retina: colocalizations with other calcium-binding proteins, GABA and glycine.

Immunocytochemical techniques were used to determine the distribution of the calcium-binding protein calretinin in the cat retina. Comparisons were made with parvalbumin and calbindin as well as with the inhibitory neurotransmitters GABA and glycine. Calretinin immunoreactivity was seen in horizontal cells and multiple subpopulations of amacrine and ganglion cells. Cone outer segments were also stained. Calbindin immunoreactivity was present in cone photoreceptors, horizontal cells, at least two subtypes of cone bipolar cell, numerous amacrine cells, and cells residing in the ganglion cell layer. Heavy staining for parvalbumin was found in both A- and B-type horizontal cells, distinct subpopulations of amacrine and ganglion cells, and a small population of cone photoreceptor cells. To confirm the identity of cone photoreceptors, comparisons were made with retinas stained for opsins specific for red/green or blue cones (Szél et al., 1986). The localization of parvalbumin corresponded with that of blue-type cones only whereas calretinin and calbindin staining showed the same distribution as both red/green and blue cones. Double-label immunofluorescence studies revealed colocalization of all three of the calcium-binding proteins in a number of neurons including horizontal cells and AII amacrine cells. To assess a possible transmitter-specific relationship for calretinin, double-label studies were carried out with GABA and glycine. However, the staining patterns for each of these inhibitory amino acids differed substantially from that of calretinin. The possibility remains that calretinin and other calcium-binding proteins may play a role in neurotransmission through interactions with receptors or second-messenger agents.

Animals↗

Gap junctions between AII amacrine cells and calbindin-positive bipolar cells in the rabbit retina.

Electrical synapses or gap junctions occur between many retinal neurons. However, in most cases, the gap junctions have not been visualized directly. Instead, their presence has been inferred from tracer spread throughout the network of cells. Thus, tracer coupling is taken as a marker for the presence of gap junctions between coupled cells. AII amacrine cells are critical interneurons in the rod pathway of the mammalian retina. Rod bipolar cell output passes to AII amacrine cells, which in turn make conventional synapses with OFF cone bipolar cells and gap junctions with ON cone bipolar cells. Injections of biotinylated tracers into AII amacrine cells reveals coupling between the AII amacrine cell network and heterologous coupling with a variety of ON cone bipolar cells, including the calbindin-positive cone bipolar cell. To directly visualize gap junctions in this network, we prepared material for electron microscopy that was double labeled with antibodies to calretinin and calbindin to label AII amacrine cells and calbindin-positive cone bipolar cells, respectively. AII amacrine cells were postsynaptic to large vesicle-laden rod bipolar terminals, as previously reported. Gap junctions were identified between AII amacrine cells and calbindin-positive cone bipolar cell terminals identified by the presence of immunostaining and ribbon synapses. This represents direct confirmation of gap junctions between two different yet positively identified cells, which are tracer coupled, and provides additional evidence that tracer coupling with Neurobiotin indicates the presence of gap junctions. These results also definitively establish the presence of gap junctions between AII amacrine cells and calbindin bipolar cells which can therefore carry rod signals to the ON alpha ganglion cell.

Animals↗

Brain calbindin-D28k and an Mr 29,000 calcium binding protein in cerebellum are different but related proteins: evidence obtained from sequence analysis by tandem mass spectrometry.

A calcium binding protein of Mr 29,000 which cross-reacts with antibodies raised against chick calbindin-D28k was previously reported to be present in rat cerebellum [Pochet, R., Parmentier, M., Lawson, D. E. M., & Pasteels, J. L. (1985) Brain Res. 345, 251-254]. It was suggested that the Mr 29,000 protein represents another form of calbindin-D28k. In our laboratory we were able to identify Mr 28,000 and 29,000 proteins in rat, human, and chick cerebellum by their ability to bind 45Ca in a 45Ca blot assay. Two calcium binding proteins of Mr 27,680 and 29,450 were isolated from rat cerebelli by the use of gel permeation chromatography and preparative gel electrophoresis. After reverse-phase high-performance liquid chromatography (HPLC) the proteins were sequenced. Sequence analysis by tandem mass spectrometry indicated only 52% identity between the rat cerebellar Mr 28,000 and 29,000 proteins. Thus they are not different forms of the same protein, as previously suggested. Eighty-nine percent identity was observed between the rate cerebellar Mr 29,000 protein and chick calretinin [Rogers, J. H. (1987) J. Cell Biol. 105, 1343-1353]. The difference in identity between the rat cerebellar Mr 29,000 protein and chick calretinin may be due to species differences, and thus this protein is most likely rat calretinin. However, barely detectable levels of mRNA and weak immunoreactivity have been reported for calretinin in cerebellum. Since we found the Mr 29,000 calcium binding protein to be an abundant protein in rat cerebellum, these results suggest either posttranscriptional regulation of calretinin in cerebellum or species differences.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Preproenkephalin-like immunoreactive and calcium-binding proteins-like immunoreactive double-labelled neurons in the spinal trigeminal nucleus caudalis of the rat.

Immunofluorescence histochemical double-staining for preproenkephalin (PPE) and calbindin-D28k (CB), calretinin (CR) or parvalbumin (PV) were performed in the spinal trigeminal nucleus caudalis (Vc) of the rat. Neuronal cell bodies exhibiting PPE-like immunoreactivity were present in all laminae of the Vc, with a higher concentration in lamina II. Most of the CB-, CR- and PV-like immunoreactive neurons were located in lamina II, and some of them were also found in laminae I and III of the Vc. Some PPE-like immunoreactive neurons also showed CB-, CR-, or PV-like immunoreactivities. CB/PPE, CR/PPE and PV/PPE double-labelled neurons were mainly observed in lamina II. The percentages of CB/PPE double-labelled neurons in the total numbers of the CB- and PPE-like immunoreactive neurons were 3.5-1.5% and 3.3-15.7%, respectively. Of all CR- and PPE-like immunoreactive neurons, 4.7-13.5% and 3.7-14.2% showed both CR- and PPE-like immunoreactivities. The ratios of PV/PPE double-labelled neurons in all PV- and PPE-like immunoreactive neurons were 9.7-28.1% and 2.1-8.7%, respectively. The present results indicate that some enkephalinergic neurons in the Vc of the rat also contain calcium-binding proteins.

Animals↗

Calretinin modifies presynaptic calcium signaling in frog saccular hair cells.

To determine whether the concentrations of calcium-binding proteins present in some neurons and sensory cells are sufficient to influence presynaptic calcium signaling, we studied the predominant calcium-binding protein in a class of sensory hair cells in the frog ear. Based on antibody affinity and molecular weight, we identified this protein as calretinin. We measured its cytoplasmic concentration to be approximately 1.2 mM, sufficient to bind approximately 6 mM Ca2+. Calcium signaling was altered when the diffusible cytoplasmic components were replaced by an intracellular solution lacking any fast calcium buffer, and was restored by the addition of 1.2 mM exogenous calretinin to the intracellular solution. We conclude that calretinin, when present at millimolar concentration, can serve as a diffusionally mobile calcium buffer/transporter capable of regulating calcium signaling over nanometer distances at presynaptic sites.

Animals↗

The neurotensin agonist PD149163 increases Fos expression in the prefrontal cortex of the rat.

Dopaminergic axons innervating the prefrontal cortex (PFC) target both pyramidal cells and GABAergic interneurons. Many of these dopamine (DA) axons in the rat coexpress the peptide neurotransmitter neurotensin. Previous electrophysiological data have suggested that neurotensin activates GABAergic interneurons in the PFC. Activation of D2-like DA receptors increases extracellular GABA levels in the PFC, as opposed to the striatum, where D2 receptor activation inhibits GABAergic neurons. Because activation of presynaptic D2 release-modulating autoreceptors in the PFC suppresses DA release but increases release of the cotransmitter neurotensin, D2 agonists may enhance the activity of GABAergic interneurons via release of neurotensin. In order to determine if neurotensin can activate GABAergic interneurons, we treated rats with the peptide neurotensin agonist, PD149163, and examined Fos expression in PFC neurons. Systemic administration of PD149163 increased overall Fos expression in the PFC, but not in the dorsal striatum. PD149163 induced Fos in PFC interneurons, as defined by the presence of calcium-binding proteins, and in pyramidal cells. Pretreatment with the high-affinity neurotensin antagonist, SR48692, blocked neurotensin agonist-induced Fos expression. These data suggest that neurotensin activates interneurons in the PFC of the rat.

Animals↗

Structural and biochemical characterization of neuronal calretinin domain I-II (residues 1-100). Comparison to homologous calbindin D28k domain I-II (residues 1-93).

This study characterizes the calcium-bound CR I-II domain (residues 1-100) of rat calretinin (CR). CR, with six EF-hand motifs, is believed to function as a neuronal intracellular calcium-buffer and/or calcium-sensor. The secondary structure of CR I-II, defined by standard NMR methods on 13C,15N-labeled protein, contains four helices and two short interacting segments of extended structure between the calcium-binding loops. The linker between the two helix-loop-helix, EF-hand motifs is 12 residues long. Limited trypsinolysis at K60 (there are 10 other K/R residues in CR I-II) confirms that the linker of CR I-II is solvent-exposed and that other potential sites are protected by regular secondary structure. 45Ca-overlay of glutathione S-transferase (GST)-CR(1-60) and GST-CR(61-100) fusion proteins confirm that both EF-hands of CR I-II have intrinsic calcium-binding properties. The primary sequence and NMR chemical shifts, including calcium-sensitive glycine residues, also suggest that both EF-hand loops of CR I-II bind calcium. NMR relaxation, analytical ultracentrifugation, chemical cross-linking and NMR translation diffusion measurements indicate that CR I-II exists as a monomer. Calb I-II (the homologous domain of calbindin D28k) has the same EF-hand secondary structures as CR I-II, except that helix B is three residues longer and the linker has only four residues [Klaus, W., Grzesiek, S., Labhardt, A. M., Buckwald, P., Hunziker, W., Gross, M. D. & Kallick, D. A. (1999) Eur. J. Biochem. 262, 933-938]. In contrast, Calb I-II binds one calcium cation per monomeric unit and exists as a dimer. Despite close homology and similar secondary structures, CR I-II and Calb I-II probably have distinct tertiary structure features that suggest different cellular functions for the full-length proteins.

Amino Acid Sequence↗