Posttranslational processing and differential glycosylation of Tractin, an Ig-superfamily member involved in regulation of axonal outgrowth

Jie, Chunfa; Xu, Yingzhi; Wang, Dong; Lukin, Dana J.; Zipser, Birgit; Jellies, John; Johansen, Kristen M.; Johansen, Jørgen

doi:10.1016/s0167-4838(00)00030-3

Cited by 11 publications

(15 citation statements)

References 55 publications

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“…1A, arrows), one in the third FNIII-domain, and one just proximal to the transmembrane domain (22). Previously, a full length version of the Tractin protein has not been observed on immunoblots of leech central nervous system proteins (21,22); however, on greatly overloaded gels the same faint band corresponding to the expected size for the unprocessed protein can be detected with different domain specific antibodies ( …”

Section: Resultsmentioning

confidence: 98%

“…It contains 6 Igdomains, 4 FNIII-like domains, an acidic domain, 12 repeats of a novel collagen-like prolineand glycine-rich sequence motif, a transmembrane domain, and an intracellular tail with an ankyrin and a PDZ-domain binding motif (21,22). The predicted molecular weight of Tractin is 198 kDa; however, antibodies to the NFb-terminal sequence of Tractin recognize only a 130 kDa glycosylated fragment on standard immunoblots (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We found that both the Laz6-56 and IH4 antibodies can coimmunoprecipitate the transmembrane as well as the cleaved NHi-terminal secreted fragment from leech nerve cord extracts strongly indicating that the two fragments also are likely to interact in the nervous system. Interestingly, the secreted middle fragment generated from proteolysis at the furin site as well as just distal to the transmembrane segment is also located to the surface of neurons and axons (22) The migration of molecular mass markers in kDa is indicated in grey. cell lysate cell medium …”

Section: Discussionmentioning

confidence: 99%

“…The previously reported Tractin monoclonal antibodies (mAbs), Laz6-56,4G5, 3A11, and 3A12 (20)(21)(22) were used in these studies. In addition a new antibody, IH4, was made to a glutathione-S-transferase (GST)-fusion protein of the PGYG-domain of Tractin in the pGEX vector system (Pharmacia) encompassing the sequence T1184-P1662.…”

Section: Antibodiesmentioning

confidence: 99%

“…IB) are much higher than the 76 and 98 kDa predicted for these bands based on their amino acid sequence. This discrepancy can either be due to homo-and/or heterodimer formation through the formation of SDS resistant covalent bonds or to anomalous gel migration (22). In order to distinguish between these possibilities we compared the migration on SDS-PAGE of Tractin constructs expressed in the S2 cell line where different COOH-terminal sequences had been deleted (Fig.…”

Section: Expression and Processing Of Tractin In The S2 Cell Linementioning

confidence: 99%

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Posttranslational processing and functional analysis of Tractin, an L1 family member in leech

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Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Antibodiesmentioning

confidence: 99%

Section: Expression and Processing Of Tractin In The S2 Cell Linementioning

confidence: 99%

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Posttranslational processing and functional analysis of Tractin, an L1 family member in leech

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In vivo dynamics of CNS sensory arbor formation: A time‐lapse study in the embryonic leech

et al. 2003

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In the embryo of the leech Hirudo medicinalis, afferent projections of peripheral sensory neurons travel along common nerve tracts to the CNS, where they defasciculate, branch, and arborize into separate, modality-specific synaptic laminae. Previous studies have shown that this process requires, at least in part, the constitutive and then modality-specific glycosylations of tractin, a leech L1 homologue. We report here on the dynamics of growth of these projections as obtained by examining the morphology of single growing dye-filled sensory afferents as a function of time. Using 2-photon laser-scanning microscopy of the intact developing embryo, we obtained images of individual sensory projections at 3 to 30 min intervals, over several hours of growth, and at different stages of development. The time-lapse series of images revealed a highly dynamic and maturation-state-dependent pattern of growth. Upon entering the CNS, the growth cone-tipped primary axon sprouted numerous long filopodial processes, many of which appeared to undergo repeated cycles of extension and retraction. The growth cone was transformed into a sensory arbor through the formation of secondary branches that extended within the ganglionic neuropil along the anterior-posterior axis of the CNS. Numerous tertiary and quaternary processes grew from these branches and also displayed cycles of extension and retraction. The motility of these higher-order branches changed with age, with younger afferents displaying higher densities and greater motility than older, more mature sensory arbors. Finally, coincident with a reduction in higher order projections was the appearance of concavolar structures on the secondary processes. Rows of these indentations suggest the formation of presynaptic en-passant specializations accompanying the developmental onset of synapse formation.

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Leech filamin and Tractin: Markers for muscle development and nerve formation

Venkitaramani

Wang

et al. 2004

J. Neurobiol.

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The Lan3-14 and Laz10-1 monoclonal antibodies recognize a 400 kDa antigen that is specifically expressed by all muscle cells in leech. We show that the antigen recognized by both antibodies is a member of the filamin family of actin binding proteins. Leech filamin has two calponin homology domains and 35 filamin/ABP-repeat domains. In addition, we used the Laz10-1 antibody to characterize the development of the segmentally iterated dorsoventral flattener muscles. We demonstrate that the dorsoventral flattener muscle develops as three discrete bundles of myofibers and that CNS axons pioneering the DP nerve extend only along the middle bundle. Interestingly, the middle dorsoventral muscle anlage is associated with only non-neuronal expression of the L1-family cell adhesion molecule Tractin. This expression is transient and occurs at the precise developmental stages when DP nerve formation takes place. Based on these findings we propose that the middle dorsoventral muscle anlagen provides a substrate for early axonal outgrowth and nerve formation and that this function may be associated with differential expression of distinct cell adhesion molecules.

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Posttranslational processing and differential glycosylation of Tractin, an Ig-superfamily member involved in regulation of axonal outgrowth

Cited by 11 publications

References 55 publications

Posttranslational processing and functional analysis of Tractin, an L1 family member in leech

Posttranslational processing and functional analysis of Tractin, an L1 family member in leech

In vivo dynamics of CNS sensory arbor formation: A time‐lapse study in the embryonic leech

Leech filamin and Tractin: Markers for muscle development and nerve formation

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