Mass spectrometry strategies applied to the characterization of proline-rich peptides from secretory parotid granules of pig (Sus scrofa).
Basic proline-rich proteins (bPRPs) are a class of proteins widely present in saliva of humans and other mammals. They are synthesized as preproproteins and enzymatically cleaved into small peptides before secretion from the salivary glands. Recently, we characterized two proline-rich peptides (SP-A and SP-B) in parotid secretory granules of pig (Sus Scrofa) that are derived from three isoforms of a PRP proprotein (Swiss-Prot data bank: Q95JC9-1, Q95JC9-2 and Q95JC9-3). Together the coding regions for SP-A and SP-B, which are repeated many times, account for 52-70% of the coding regions of the PRP proproteins. This study was undertaken to identify peptides encoded by unassigned regions of the PRP proproteins. RP-HPLC-ESI-IT-MS analysis of enriched granule preparations from pig parotid glands by two different analytical strategies identified ten new proline-rich peptides derived from the three proproteins. Together with the coding regions for SP-A and SP-B already identified it was possible to assign 68-75% of the proproteins coding regions. The peptide sequences indicated a number of unusual proteolytic cleavage sites suggesting the presence of unknown proprotein convertases.
Fanali
Chiara
C
Institute of Biochemistry and Clinical Biochemistry, Catholic University, Largo F. Vito, Rome, Italy.
Inzitari
Rosanna
R
Cabras
Tiziana
T
Fiorita
Antonella
A
Scarano
Emanuele
E
Patamia
Maria
M
Retruzzelli
Raffaele
R
Bennick
Anders
A
Messana
Irene
I
Castagnola
Massimo
M
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010306:Parotid Gland / Q000737:chemistry*; D010455:Peptides / Q000032:analysis*; D055232:Proline-Rich Protein Domains; D020033:Protein Isoforms / Q000032:analysis; D015203:Reproducibility of Results; D022142:Secretory Vesicles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013552:Swine
D010455:Peptides; D020033:Protein Isoforms