Proteomic Analysis of a 14.2 kDa Protein Isolated from Bali Cattle(Bos Sondaicus/Javanicus)Saliva Using Single Dimension Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis and Matrix-Assisted Laser Desorption Ionization Time-of-Flight Tandem Mass Spectrometer

Abstract

A 14.2 kDa protein isolated from Bali cattle (Bos sondaicus/javanicus) saliva has been reported to have a bactericidal activity. The aim of this study was to analyse the nature of a 14.2 kDa protein using single dimension (1-D) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometer (MALDI TOF/TOF). The protein was isolated by means of polyethylene glycol (PEG)/sodium sulfate aqueous twophase system, and then determined by 12.5% SDS-PAGE. A band of 14.2 kDa was sliced and analysed by MALDI TOF/TOF mass spectrometer using a 5800 proteomics analyzer. Mascot search and National Center for Biotechnology Information (NCBI) Blast search revealed that the spot of 1-D SDS-PAGE consisted of three proteins: zymogen granule protein 16 homologue B, pancreatic adenocarcinoma upregulated factor-like, and prolactin-inducible protein homologue precursor. The three proteins have been observed in Bos taurus and other species such as mouse. The actual nature of the proteins and their function in Bali cattle (Bos sondaicus/javanicus), as well as the connection with the evolution of bovines need further analysis.

The Utilization of the SDS-PAGE Salivary Protein Profiles for Clustering Analysis of Bali Cattle (Bos sondaicus/javanicus) and the Taurine or Zebu Cross Breeds in North Lombok District, Indonesia: A Preliminary Study

“…Their coat color is very distinctive, usually reddishbrown, except for a clearly defined white area on the hindquarters that extends along the belly, and also white socks reaching from the hooves to just above the hocks compiled by Lindell [9]. Saliva was collected and treated as described previously [10] from oral cavity of the cattle, using disposable plastic pipettes. About 5-10 ml of saliva collected in disposable glass tubes containing preservative/ bactericidal (0.2% v/v NaN 3 ) were placed in a cool box containing ice and immediately taken to the laboratory for further treatments.…”

“…In this study salivary protein between the species in the region of 17-45 kDa molecular weights were preferred for analyzing due to some specific markers for bovine saliva that have been found in that range [10,[13][14][15]. Only the unambiguous bands were scored or coded for the presence (scored 1) or absence (scored 0) of each saliva sample.…”

Proteomics and sequence analysis of a 30-kDa saliva protein of Indonesian local goat (Capra hircus) using MALDI-TOF/TOF-MS and related techniques

“…Whole saliva was collected from the oral cavity of 50 healthy male and female Indonesian local goats (age range: 4-30 months) using a plastic pipette, based on a previously published method with some modifications (15). All animals used in this study were kept under similar conditions.…”

“…Identification of the 30-kDa protein was carried out according to a previous study (15). A stained band that corresponded to a 30-kDa relative mass was excised and sent to Proteomics International Pty.…”

Complement C3 and anti-testosterone antibody in the 150 kDa Bali cattle (Bos sondaicus/javanicus) subordinate follicular fluid peptide observed using MALDI-TOF/TOF-MS

“…Briefly the samples were reduced, alkylated and trypsin digested, extracted and then analyzed by MALDI-TOF/TOF-MS using a 5800 Proteomics Analyzer (AB Sciex, Framngham, MA, USA). In order to identify the protein of interest, spectra obtained were analyzed using Mascot sequence matching software (Matrix Science, Boston, MA, USA) with Ludwig NR Database, as performed by Depamede (2013).…”