Protein profiling of water and alkali soluble cottonseed protein isolates

Abstract

Currently, there is only limited knowledge on the protein types and structures of the cottonseed proteins. In this work, water-soluble cottonseed proteins (CSPw) and alkali-soluble cottonseed proteins (CSPa) were sequentially extracted from defatted cottonseed meal. Proteins of the two fractions were separated by 4-20% gradient polyacrylamide gel electrophoresis (SDS-PAGE); There were 7 and 12 polypeptide bands on SDS-PAGE of CSPa and CSPw, respectively. These individual bands were then excised from the gel and subjected to mass spectrometric analysis. There were total 70 polypeptides identified from the proteins of the two cottonseed preparations, with molecular weights ranging from 10 to 381 kDa. While many proteins or their fragments were found in multiple bands, 18 proteins appeared only in one SDS-PAGE band (6 in CSPa, 12 in CSPw). Putative functions of these proteins include storage, transcription/translation, synthesis, energy metabolism, antimicrobial activity, and embryogenesis. Among the most abundant are legumin A (58 kDa), legumin B (59 kDa), vicilin C72 (70 kDa), vicilin GC72-A (71 kDa), and vicilin-like antimicrobial peptides (62 kDa). This work enriched the fundamental knowledge on cottonseed protein composition, and would help in better understanding of the functional and physicochemical properties of cottonseed protein and for enhancing its biotechnological utilization.

Figure 1

Gradient (4–12%) SDS-PAGE of two-step prepared water- (CSPw, left) and alkali- (CSPa, right) soluble cottonseed protein isolates. Approximately 5 µg of protein were applied to each lane).

Figure 2

GO term predicted biological functions and percentage of each class of proteins identified in cottonseeds.

Figure 3

Sequence coverage of 40 S ribonsomal S5 (A0A0B0PCC1, G. arboreum) by peptide fragments in CSPw and CSPa.

Figure 4

Sequence coverage of late embryogenesis abundant protein D-19 (P09943, G. hirsutum) by peptide fragments in CSPw and CSPa.

Figure 5

Sequence coverage of Elongation factor 1-alpha (A0A0B0P186, G. arboreum) by peptide fragments in CSPa.

Figure 6

Sequence coverage of Vicilin C72 (P09801, G. hirsutum) by peptide fragments in CSPw and CSPa.