Primary structure and possible origin of the non-glycosylated basic proline-rich protein of human submandibular/sublingual saliva

Robinson, Ranga; Kauffman, Dorothy L.; Waye, Mary Miu Yee; Blum, Max; Bennick, Anders; Keller, Patricia J.

doi:10.1042/bj2630497

Cited by 30 publications

(29 citation statements)

References 36 publications

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“…6). The binding of ECA, which reacts well with terminal Gal␤1-4GlcNAc and Fuc␣1-2Gal␤1-4GlcNAc (28,29), was abolished in cultures of S. gordonii, S. oralis, and S. mitis, while the binding of other lectins to different features of Nlinked glycan chains shifted from the ϳ70-kDa region in control saliva down to the region from ϳ35 to 60 kDa, which includes the regions (35 to 46 kDa) expected for PRB3 and PRB4 protein chains (30). N-Acetylglucosamine with an ␣-1-6-linked fucose branch (31) is the innermost (peptide-linked) monosaccharide of PRG.…”

Section: Discussionmentioning

confidence: 98%

Differential Utilization of Basic Proline-Rich Glycoproteins during Growth of Oral Bacteria in Saliva

Zhou

Yang

Zhang

et al. 2016

Appl Environ Microbiol

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Although saliva is widely recognized as a primary source of carbon and nitrogen for growth of the dental plaque biofilm community, little is known about how different oral bacteria utilize specific salivary components. To address this question, 32 strains representing 16 genera commonly isolated from early plaque biofilms were compared for growth over two transfers in stimulated (by chewing Parafilm) whole saliva that was stabilized by heat treatment and dialysis. The cell densities, measured by quantitative PCR (qPCR), ranged from ϳ1 ؋ 10 6 to 1 ؋ 10 7 /ml for strains of Streptococcus gordonii, Streptococcus oralis, and Streptococcus mitis and one strain of Streptococcus sanguinis. Strains of Streptococcus mutans, Gemella haemolysans, and Granulicatella adiacens reached ϳ1 ؋ 10 5 to 1 ؋ 10 6 /ml. In contrast, little or no growth was noted for three other strains of S. sanguinis, as well as for strains of Streptococcus parasanguinis, Streptococcus salivarius, Streptococcus vestibularis, Streptococcus sobrinus, Actinomyces spp., Abiotrophia defectiva, and Rothia dentocariosa. SDS-PAGE, lectin blotting, and two-dimensional gel electrophoresis of saliva from cultures of S. gordonii, S. oralis, and S. mitis revealed species-specific differences in the degradation of basic proline-rich glycoproteins (PRG). In contrast, saliva from cultures of other bacteria was indistinguishable from control saliva. Species-dependent differences in the utilization of individual host sugars were minor. Thus, differences in salivary glycan foraging between oral species may be important to cross-feeding and cooperation between organisms in dental plaque biofilm development. IMPORTANCEBacteria in the mouth use saliva for nutrition. How each of the many types of bacteria uses saliva is not clear. We show that a major protein in saliva, called PRG, is an important nutrition source for certain bacteria but not for others. PRG has many sugar molecules linked in chains, but the sugar is not available for bacteria until the chains are degraded. The bacteria that can grow by digesting this protein break the sugar chains into parts which not only support their own growth but could also be available to support the growth of those bacteria that cannot use the intact protein. Dental plaque biofilm development involves an array of specific interactions between bacteria and saliva, beginning with the attachment of early colonizers, including Streptococcus sanguinis, Streptococcus oralis, Streptococcus gordonii, and Streptococcus mitis, through binding to salivary components in the acquired enamel pellicle (1, 2). While interbacterial adhesion (i.e., coaggregation) between these pioneers and other bacteria enhances species diversity in the nascent biofilm (3), community maturation depends primarily on the growth of different closely associated bacteria (4, 5). Growth of bacteria, measured as an increase in optical density (OD), was noted in saliva inoculated with dental plaque (6) but not in saliva inoculated with monocultures of Streptococcus...

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Section: Discussionmentioning

confidence: 98%

Differential Utilization of Basic Proline-Rich Glycoproteins during Growth of Oral Bacteria in Saliva

Zhou

Yang

Zhang

et al. 2016

Appl Environ Microbiol

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“…and this may explain why some of the secreted proteins have higher molecular weights than those synthesized in vitro. Some human PRPs are cleaved after synthesis (Robinson et al, 1989), and similar processes may occur in rabbit parotid gland, thereby at least partly explaining how a small number of precursor proteins can give rise to a large number of secreted proteins. The rabbit basic PRPs share immunological characteristics with the proline-rich repeat region of human PRPs (Ty region) since antibodies to Ty and human PRPs give almost identical immunoblot patterns ( Figures IC and 1E).…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, the prolinepoor E region, which is not present in most human basic PRPs, is also absent in rabbit basic PRPs ( Figure 1D). At the level of stringency used in Northern blotting the HaeIII probe, which corresponds to the repeat region of human acidic PRPs, and the BstNl probe, which encodes the repeat found in human basic PRPs, will both hybridize with mRNA for human acidic and basic PRPs (Robinson et al, 1989). Since rabbit basic PRPs crossreact with antibodies to human PRPs but rabbit acidic PRPs do not (Spielman et al, 1991), it is likely that the hybridization of rabbit parotid RNA with the human cDNA probes reflects the presence of mRNA for rabbit basic PRPs.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, it has been found that in humans the distribution of individual members of the PRP family varies in the different salivary glands. Whereas acidic and basic PRPs both are found in the parotid gland, only acidic PRPs are present in the submandibular gland (Robinson et al, 1989). In mice, isoproterenol induces different patterns of PRP synthesis in the parotid and submandibular glands (Mehansho et al, 1985).…”

Section: (J T T I S T D a Cytochem 40:1393-14m 1992)mentioning

confidence: 99%

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Differential expression of proline-rich proteins in rabbit salivary glands.

Ferreira

Robinson

Hand³

et al. 1992

J Histochem Cytochem.

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Salivary glands synthesize and secrete an unusual family of proline-rich proteins (PRPs) that can be broadly divided into acidic and basic PRPs. We studied the tissue-specific expression of these proteins in rabbits, using antibodies to rabbit acidic and basic PRPs as well as antibodies and cDNA probes to human PRPs. By immunoblotting, in vitro translation, and Northern blotting, basic PRPs could be readily detected in the parotid gland but were absent in other salivary glands. In contrast, synthesis in vitro of acidic PRPs was detected in parotid, sublingual, and submandibular glands. Ultrastructural localization with immunogold showed heavy labeling with antibodies to acidic PRPs of secretory granules of parotid acinar cells and sublingual serous demilune cells. Less intense labeling occurred in the seromucous acinar cells of the submandibular gland. With antibodies to basic PRPs, the labeling of the parotid gland was similar to that observed with antibodies to acidic PRPs, but there was only weak labeling of granules of a few sublingual demilune cells, and no labeling of the submandibular gland. These results demonstrate a variable pattern of distribution of acidic and basic PRPs in rabbit salivary glands. These animals are therefore well suited for study of differential tissue expression of PRPs.

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“…The C-terminal 14-kD fragment of serum HRG is exceptionally rich in protein content. The possible relationship of this portion of HRG to the proline-rich polypeptide also found in milk is intriguing, especially given the reported immunemodulating activities attributed to proline-rich polypeptide (38)(39)(40)(41)(42). Given the recent implication that HRG plays a role in regulating T-lymphocyte activation (35,43), it may be important to elucidate both the structural and functional integrity of HRG in milk, as well as its metabolic fate in infants fed human milk.…”

Section: Discussionmentioning

confidence: 99%

Identification of Histidine-Rich Glycoprotein in Human Colostrum and Milk

1992

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ABSTRACT. Histidine-rich glycoprotein (HRG) is a 74-kD glycoprotein, originally discovered in plasma, which contains an unusually large amount of histidine (13 mol%) and proline (13 mol%). The specific functions of this protein remain unclear, although it binds (reversibly) transition metal ions such as Cu(I1) and Zn(1I) with high capacity (10-13 equivalent) and moderate to high affinity (h = 0.2-10 pM). Because the bioavailability of Cu (I1) and Zn(I1) ions in human milk is high, we have used specific antibodies from polyclonal antisera directed against purified human plasma HRG to investigate whether this or a related protein is a component of human colostrum and(or) mature milk. Fresh human colostrum (d 1-3) and milk (d 4-120) were collected in the presence and absence of multiple protease inhibitors and EDTA. Immuno "dot" blot analyses and ELISA were developed; HRG was present in both colostrum (0.13-10 pg/mL) and mature milk (0.1-10 pg/mL). Unidentified components in colostrum and milk, however, were found to depress HRG antigenicity in these assays. Western transfer and immunoblots of denatured colostrum and milk samples analyzed by SDS-PAGE revealed the presence of an immunoreactive band at 74-78 kD, with other bands at 47 and 24 kD under both reducing and nonreducing conditions; smaller immunoreactive fragments (12-14 kD) were detected in some samples. We observed at least one additional band of immunoreactivity of greater molecular mass (>I10 kD) in colostrum under nonreducing conditions; we did not observe these bands in plasma samples. Immunoaffinity and Zn(I1) affinity isolation of HRG from colostrum and milk resulted in the copurification of several associated proteins. Nterminal amino acid sequence analysis of the isolated 74-kD immunoreactive protein provided further evidence for identification as intact HRG; 13 of the first 15 residues were identical. These results demonstrate the presence of HRG, or a structurally related protein, in human colostrum and milk. Understanding the biology and function of trace elements in human. milk must include the identification of specific human milk peptides and proteins that influence metal ion transport (bioavailability) and storage in the mammary gland during lactogenesis, as well as in the gastrointestinal tract of breast-fed infants. Equally important, in our view, is the need to understand the role of transition metal ions in the regulation of milk protein structure, bioavailability, and function. Much can be learned from existing knowledge of the transport and use of transition metal ions by serum proteins. For example, in several species, including humans, many of the proteins in maternal serum, particularly the metal-binding proteins, are similar or identical to the whey proteins in colostrum (1,2).Adult human serum contains a protein discovered in 1972 (3) referred to as histidine-rich glycoprotein.' HRG is a 74-kD glycoprotein that contains significant quantities of both histidine (13%) and proline (13%) (4), which are concentrated primarily in the C...

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Primary structure and possible origin of the non-glycosylated basic proline-rich protein of human submandibular/sublingual saliva

Cited by 30 publications

References 36 publications

Differential Utilization of Basic Proline-Rich Glycoproteins during Growth of Oral Bacteria in Saliva

Differential Utilization of Basic Proline-Rich Glycoproteins during Growth of Oral Bacteria in Saliva

Differential expression of proline-rich proteins in rabbit salivary glands.

Identification of Histidine-Rich Glycoprotein in Human Colostrum and Milk

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