Purification and characterization of a small (7.3 kDa) putative lipid transfer protein from maize seeds

Castro, Mariana S.; Gerhardt, Isabel Rodrigues; Orrú, Stefania; Pucci, Piero; Bloch, Carlos

doi:10.1016/s1570-0232(03)00423-9

Cited by 30 publications

(18 citation statements)

References 33 publications

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“…LTPs are mainly grouped into two subfamilies: LTP1 members have 90 to 95 amino acid residues and are approximately 10 kD, while LTP2 members have approximately 70 amino acid residues and are 7 kD (Carvalho and Gomes, 2007). Both LTP1 and LTP2 family members are basic polypeptides with pI values from 8.5 to 12 and have a characteristic eight-Cys motif, forming four disulfide bonds, contributing to their binding activity for different lipids and hydrophobic compounds in vitro (Douliez et al, 2000;Liu et al, 2002;Castro et al, 2003;Carvalho and Gomes, 2007). In addition, LTPs contain a signal peptide at the N terminus, generally 21 to 27 amino acids for the LTP1 family and 27 to 35 amino acids for the LTP2 family (Carvalho and Gomes, 2007), allowing mature LTP peptides to enter the secretory pathway.…”

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confidence: 99%

OsC6, Encoding a Lipid Transfer Protein, Is Required for Postmeiotic Anther Development In Rice

et al. 2010

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Synthesis of lipidic components in anthers, including of the pollen exine, is essential for plant male reproductive development. Plant lipid transfer proteins (LTPs) are small, abundant lipid-binding proteins that have the ability to exchange lipids between membranes in vitro. However, their biological role in male reproductive development remains less understood. Here, we report the crucial role of OsC6 in regulating postmeiotic anther development in rice (Oryza sativa). Found in monocots, OsC6 belongs to a distinct clade from previously identified LTP1 and LTP2 family members found in both dicots and monocots. OsC6 expression is mainly detectable in tapetal cells and weakly in microspores from stage 9 to stage 11 of anther development. Immunological assays indicated that OsC6 is widely distributed in anther tissues such as the tapetal cytoplasm, the extracellular space between the tapetum and middle layer, and the anther locule and anther cuticle. Biochemical assays indicated that recombinant OsC6 has lipid binding activity. Moreover, plants in which OsC6 was silenced had defective development of orbicules (i.e. Ubisch bodies) and pollen exine and had reduced pollen fertility. Furthermore, additional evidence is provided that the expression of OsC6 is positively regulated by a basic helix-loop-helix transcription factor, Tapetum Degeneration Retardation (TDR). Extra granule-like structures were observed on the inner surface of the tdr tapetal layer when the expression of OsC6 was driven by the TDR promoter compared with the tdr mutant. These data suggest that OsC6 plays a crucial role in the development of lipidic orbicules and pollen exine during anther development in rice.

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confidence: 99%

OsC6, Encoding a Lipid Transfer Protein, Is Required for Postmeiotic Anther Development In Rice

et al. 2010

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“…Plant nsLTPs have been classified into two groups: type 1 (LTP1, ϳ10 kDa) and type 2 (LTP2, ϳ7 kDa) (10,50). Plant nsLTPs are often purified as several isoforms with very similar sequences (52,(55)(56)(57). To date, no one FIGURE 7.…”

Section: Discussionmentioning

confidence: 99%

Two SCA (Stigma/Style Cysteine-rich Adhesin) Isoforms Show Structural Differences That Correlate with Their Levels of in Vitro Pollen Tube Adhesion Activity

Chae

Zhang

et al. 2007

Journal of Biological Chemistry

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Lily pollen tubes grow adhering to an extracellular matrix produced by the transmitting tract epidermis in a hollow style. SCA, a small (ϳ9.4 kDa), basic protein plus low esterified pectin from this extracellular matrix are involved in the pollen tube adhesion event. The mode of action for this adhesion event is unknown. We partially separated three SCA isoforms from the lily stigma in serial size exclusion column fractions (SCA1, 9370 Da; SCA2, 9384 Da; SCA3, 9484 Da). Peptide sequencing analysis allowed us to determine two amino acid variations in SCA3, compared with SCA1. For SCA2, however, there are more sequence variations yet to be identified. Our structural homology and molecular dynamics modeling results show that SCA isoforms have the plant nonspecific lipid transfer protein-like structure: a globular shape of the orthogonal 4-helix bundle architecture, four disulfide bonds, an internal hydrophobic and solvent-inaccessible cavity, and a long C-terminal tail. The Ala 71 in SCA3, replacing the Gly 71 in SCA1, has no predictable effect on structure. The Arg 26 in SCA3, replacing the Gly 26 in SCA1, is predicted to cause structural changes that result in a significantly reduced volume for the internal hydrophobic cavity in SCA3. The volume of the internal cavity fluctuates slightly during the molecular dynamics simulation, but overall, SCA1 displays a larger cavity than SCA3. SCA1 displays higher activity than SCA3 in the in vitro pollen tube adhesion assay. No differences were found between the two SCAs in a binding assay with pectin. The larger size of the hydrophobic cavity in SCA1 correlates with its higher adhesion activity.During lily pollination, pollen tubes are guided to the ovary on an extracellular matrix (ECM) 2 produced by the specialized stylar transmitting tract epidermis (TTE). This guidance includes an adhesion event between the pollen tube and the TTE (1-4). For the purpose of identifying molecular factors in lily pollen tube adhesion, Jauh et al. (5) developed an in vitro bioassay using ECM molecules attached to a nitrocellulose membrane as an artificial style. Using this in vitro bioassay, two molecules from the style ECM, SCA (stigma/style cysteine-rich adhesin) and a low esterified pectic polysaccharide, were found to be responsible for pollen tube adhesion on the stylar matrix (6, 7). The level of SCA mRNA was high in stigma/style tissue, petal, and young leaf, but it was absent in pollen tubes (8). Localization of both SCA and the low esterified pectin on the surface of the stylar TTE supported their involvement in in vivo pollen tube adhesion. The mechanism of action for SCA and pectin in pollen tube adhesion is yet unknown. Neither of these molecules alone is active in the adhesion assay. Only when they were combined did SCA and pectin bind to each other in a pH-dependent manner and form the functional, adhesive matrix (6). The pH effect implies that a charge interaction occurs, rather than a tight cross-linking, between SCA and the low esterified pectin in lily pistils.SCA is a sm...

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“…Thus, the nsLTPs are very stable to heat and different denaturing agents (Lindorff-Larsen and Winther, 2001). Further, the nsLTP proteins are soluble and usually have a high pI due to many charged amino acids Castro et al, 2003).…”

Section: Characteristics and Structurementioning

confidence: 99%

Plant lipid transfer proteins: Evolution, expression and function

Edstam¹

2013

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Purification and characterization of a small (7.3 kDa) putative lipid transfer protein from maize seeds

Cited by 30 publications

References 33 publications

OsC6, Encoding a Lipid Transfer Protein, Is Required for Postmeiotic Anther Development In Rice

OsC6, Encoding a Lipid Transfer Protein, Is Required for Postmeiotic Anther Development In Rice

Two SCA (Stigma/Style Cysteine-rich Adhesin) Isoforms Show Structural Differences That Correlate with Their Levels of in Vitro Pollen Tube Adhesion Activity

Plant lipid transfer proteins: Evolution, expression and function

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