Crystal structure of the 30 K protein from the silkworm Bombyx mori reveals a new member of the β-trefoil superfamily

Jie-pin, Yang; Ma, Xiao-Xiao; He, Yong‐Xing; Li, Weifang; Kang, Yilan; Bao, Rui; Chen, Yuxing; Zhou, Cong‐Zhao

doi:10.1016/j.jsb.2011.04.003

Cited by 30 publications

(31 citation statements)

References 41 publications

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“…A total of 23 Kac sites were identified on 11 30 K proteins (Supporting Information Table 1). In addition to their N-terminal lipid-binding domain, 30 K proteins also contain a C-terminal sugar-binding domain [64], revealing a storage, and transport activity for carbohydrate. In addition to 30 K proteins, SPs also contain 4.8-6.9% carbohydrate [65]; whereas vitellogeninis are lipoglycoproteins with 7-16% lipid and 1-14% carbohydrate [45], suggesting that these nutrient-storage proteins may provide not only amino acids and lipids but also essential carbohydrates for development and metamorphosis.…”

Section: Lysine Acetylation May Represent a Common Regulatory Mechanimentioning

confidence: 99%

Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori

et al. 2015

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Lysine acetylation in proteins is a dynamic and reversible PTM and plays an important role in diverse cellular processes. In this study, using lysine-acetylation (Kac) peptide enrichment coupled with nano HPLC/MS/MS, we initially identified the acetylome in the silkworms. Overall, a total of 342 acetylated proteins with 667 Kac sites were identified in silkworm. Sequence motifs analysis around Kac sites revealed an enrichment of Y, F, and H in the +1 position, and F was also enriched in the +2 and -2 positions, indicating the presences of preferred amino acids around Kac sites in the silkworm. Functional analysis showed the acetylated proteins were primarily involved in some specific biological processes. Furthermore, lots of nutrient-storage proteins, such as apolipophorin, vitellogenin, storage proteins, and 30 K proteins, were highly acetylated, indicating lysine acetylation may represent a common regulatory mechanism of nutrient utilization in the silkworm. Interestingly, Ser2 proteins, the coating proteins of larval silk, were found to contain many Kac sites, suggesting lysine acetylation may be involved in the regulation of larval silk synthesis. This study is the first to identify the acetylome in a lepidoptera insect, and expands greatly the catalog of lysine acetylation substrates and sites in insects.

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Section: Lysine Acetylation May Represent a Common Regulatory Mechanimentioning

confidence: 99%

Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori

et al. 2015

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“…The β-strands forming the trefoil are connected by an intricate system of loops, all of which are well-defined in electron density. A detailed comparison of the NTD and CTD domains of the 30-kDa LPs with other structures available in the PDB was presented previously [33], [34].…”

Section: Resultsmentioning

confidence: 99%

“…To-date, only two 30-kDa LPs were characterized structurally, Bmlp7 [33], [34] and Bmlp3 [35]. In this paper we also present a detailed comparative structural analysis of the three 30-kDa LPs, and discuss how the structural differences could be related to diverse physiological roles of the members of this large protein family.…”

Section: Introductionmentioning

confidence: 99%

Crystal Structure of Bombyx mori Lipoprotein 6: Comparative Structural Analysis of the 30-kDa Lipoprotein Family

et al. 2014

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The 30-kDa lipoprotein (LP) family of mulberry silkworm comprises major hemolymph proteins specific to the fifth instar larvae. The family consists of 46 members, 24 of which are referred to as typical 30-kDa LPs. To date, two crystal structures of 30-kDa LPs from Bombyx mori have been described (Bmlp3 and Bmlp7). Here, we present the crystal structure of Bmlp6, another 30-kDa LP member. Bmlp6 is comprised of two domains characteristic of this family, the VHS-type N-terminal domain and β-trefoil C-terminal domain. The structures of the three 30-kDa LPs have been compared and a number of differences are noted, including loop conformation, the surface electrostatic potential, and the potential binding cavities. We discuss the observed structural differences in the light of the potential different roles of the particular 30-kDa LP members in silkworm physiology.

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“…They were identified as lipoproteins [25]. Yang et al [37] revealed that FB Bmlp7 plays a crucial role in the protection of B. mori against invading pathogens due to the presence of a putative sugar binding domain. Crystal structures of 30 kDa proteins LP2 (P09335) [38] and Bmlp7 [26,37] indicated an N-terminal lipid-binding cavity and a C-terminal sugar-binding site.…”

Section: Introductionmentioning

confidence: 99%

“…Yang et al [37] revealed that FB Bmlp7 plays a crucial role in the protection of B. mori against invading pathogens due to the presence of a putative sugar binding domain. Crystal structures of 30 kDa proteins LP2 (P09335) [38] and Bmlp7 [26,37] indicated an N-terminal lipid-binding cavity and a C-terminal sugar-binding site. The 30 kDa protein 6 G1 (Bmlp1) from FB was reported to interfere with hyphal growth of the entomopathogenic fungus, Paecilomyces tenuipes [35].…”

Section: Introductionmentioning

confidence: 99%

A proteomic view on the developmental transfer of homologous 30 kDa lipoproteins from peripheral fat body to perivisceral fat body via hemolymph in silkworm, Bombyx mori

et al. 2012

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BackgroundA group of abundant proteins of ~30 kDa is synthesized in silkworm larval peripheral fat body (PPFB) tissues and transported into the open circulatory system (hemolymph) in a time-depended fashion to be eventually stored as granules in the pupal perivisceral fat body (PVFB) tissues for adult development during the non-feeding stage. These proteins have been shown to act anti-apoptotic besides being assigned roles in embryogenesis and defense. However, detailed protein structural information for individual PPFB and PVFB tissues during larval and pupal developmental stages is still missing. Gel electrophoresis and chromatography were used to separate the 30 kDa proteins from both PPFB and PVFB as well as hemolymph total proteomes. Mass spectrometry (MS) was employed to elucidate individual protein sequences. Furthermore, 30 kDa proteins were purified and biochemically characterized.ResultsOne- and two-dimensional gel electrophoresis (1/2D-PAGE) was used to visualize the relative changes of abundance of the 30 kDa proteins in PPFB and PVFB as well as hemolymph from day 1 of V instar larval stage to day 6 of pupal stage. Their concentrations were markedly increased in hemolymph and PVFB up to the first two days of pupal development and these proteins were consumed during development of the adult insect. Typically, three protein bands were observed (~29, 30, 31 kDa) in 1D-PAGE, which were subjected to MS-based protein identification along with spots excised from 2D-gels run for those proteomes. Gas phase fragmentation was used to generate peptide sequence information, which was matched to the available nucleotide data pool of more than ten highly homologous insect 30 kDa lipoproteins. Phylogenetic and similarity analyses of those sequences were performed to assist in the assignment of experimentally identified peptides to known sequences. Lipoproteins LP1 to LP5 and L301/302 could be matched to peptides extracted from all bands suggesting the presence of full length and truncated or modified protein forms in all of them. The individual variants could not be easily separated by classical means of purification such as 2D-PAGE because of their high similarity. They even seemed to aggregate as was indicated by native gel electrophoresis. Multistep chromatographic procedures eventually allowed purification of an LP3-like protein. The protein responded to lipoprotein-specific staining.ConclusionsIn B. mori larvae and pupae, 30 kDa lipoproteins LP1 to LP5 and L301/302 were detected in PPFB and PVFB tissue as well as in hemolymph. The concentration of these proteins changed progressively during development from their synthesis in PPFB, transport in hemolymph to storage in PVFB. While the 30 kDa proteins could be reproducibly separated in three bands electrophoretically, the exact nature of the individual protein forms present in those bands remained partially ambiguous. The amino acid sequences of all known 30 kDa proteins showed very high homology. High-resolution separation techniques will be necessary before MS a...

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Crystal structure of the 30 K protein from the silkworm Bombyx mori reveals a new member of the β-trefoil superfamily

Cited by 30 publications

References 41 publications

Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori

Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori

Crystal Structure of Bombyx mori Lipoprotein 6: Comparative Structural Analysis of the 30-kDa Lipoprotein Family

A proteomic view on the developmental transfer of homologous 30 kDa lipoproteins from peripheral fat body to perivisceral fat body via hemolymph in silkworm, Bombyx mori

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