The hirudin-like factors HLF3 and HLF4—hidden hirudins of European medicinal leeches

Müller, Christian; Lukas, Phil; Sponholz, Dana; Hildebrandt, Jan‐Peter

doi:10.1007/s00436-020-06697-1

Cited by 12 publications

(11 citation statements)

References 28 publications

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“…Slight modifications within the N-termini converted HLF1 variants that initially had no measurable thrombin-inhibitory potency into very strong thrombin inhibitors (Müller et al 2020a). An exchange of the central globular domains of HLF2, HLF3l, and HLF4b with the central globular domain of HLF1 had the same effect (Müller et al 2020b). Similar investigations on the HLFs of Hirudinaria manillensis are an excellent opportunity to figure out whether or not the observed structure-function relationships are unique to HLFs of the genus Hirudo or can be observed in HLFs of other genera as well and may hence represent a more general principle.…”

Section: Introductionmentioning

confidence: 70%

“…The comparably high thrombin-inhibitory potency of HLF-Hyb3s was a somehow surprising observation. HLF-Hyb3s was as effective in thrombin inhibition as its direct counterpart HLF-Hyb3b despite the much shorter C-terminal tail (Müller et al 2020b). This raises the question whether HLF-Hyb3s (and HLF-Hyb3b as well) is still a bi-or rather a mono-(or uni-) valent inhibitor of thrombin.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently neither the N-termini nor the central globular domains of the respective factors are altered, but the lengths and the amino acid composition of their C-terminal tails are different. In case of HLF4, the C-terminal tail of HLF4b conveyed thrombin-inhibitory potency to the molecule, whereas the tail of HLF4a did not (Müller et al 2020b). A similar different splice event with alternative products may be predicted for the HLF6 gene, too (GenBank Acc.…”

Section: Introductionmentioning

confidence: 85%

“…Some HLF genes may encode different mRNAs (and hence different HLF molecules) due to alternative splice events. For HLF3l/HLF3s such a mechanism could already be confirmed (Müller et al 2016), and for HLF4a/HLF4b, it is likely (Müller et al 2017(Müller et al , 2020b. In either case, the different splice events affect the junction between the third and the fourth exon.…”

Section: Introductionmentioning

confidence: 90%

“…2a). In a previous study, we were able to successfully express and purify HLF3s, but the factor did not inhibit hirudin (Müller et al 2020b). As shown in Fig.…”

Section: Minimum Length Of the C-terminal Tailmentioning

confidence: 96%

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Short tail stories: the hirudin-like factors HLF6 and HLF7 of the Asian medicinal leech, Hirudinaria manillensis

et al. 2021

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The leech-derived hirudins and hirudin-like factors (HLFs) share a common molecule structure: a short N-terminus, a central globular domain, and an elongated C-terminal tail. All parts are important for function. HLF6 and HLF7 were identified in the Asian medicinal leech, Hirudinaria manillensis. The genes of both factors encode putative splice variants that differ in length and composition of their respective C-terminal tails. In either case, the tails are considerably shorter compared to hirudins. Here we describe the functional analyses of the natural splice variants and of synthetic variants that comprise an altered N-terminus and/or a modified central globular domain. All natural splice variants of HLF6 and HLF7 display no detectable thrombin-inhibitory potency. In contrast, some synthetic variants effectively inhibit thrombin, even with tails as short as six amino acid residues in length. Our data indicate that size and composition of the C-terminal tail of hirudins and HLFs can vary in a great extent, yet the full protein may still retain the ability to inhibit thrombin.

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Section: Introductionmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 85%

Section: Introductionmentioning

confidence: 90%

“…2a). In a previous study, we were able to successfully express and purify HLF3s, but the factor did not inhibit hirudin (Müller et al 2020b). As shown in Fig.…”

Section: Minimum Length Of the C-terminal Tailmentioning

confidence: 96%

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Short tail stories: the hirudin-like factors HLF6 and HLF7 of the Asian medicinal leech, Hirudinaria manillensis

et al. 2021

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Identification and functional characterization of multiple haemadins and an oligomeric decorsin in the Asian land leech Haemadipsa interrupta

Müller,

Sponholz,

Tolksdorf

et al. 2024

Parasitol Res

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Haematophagous leeches rely on a broad variety of bioactive compounds to secure a sufficient blood meal from their vertebrate prey. Both the primary (platelet aggregation) and secondary (blood coagulation) haemostasis are the main targets of action. The platelet aggregation inhibitor decorsin was first described in the North American leech, Macrobdella decora Say, 1824, whereas the bivalent thrombin inhibitor hirudin was originally identified in the European medicinal leech, Hirudo medicinalis Linnaeus, 1758. Hirudin blocks both the catalytic site and the fibrinogen-binding site (exosite I) of thrombin. Haemadin of the Indian land leech Haemadipsa sylvestris Blanchard, 1894, is also a highly efficient bivalent thrombin inhibitor but blocks exosite II of thrombin. So far, only the archetypal form of haemadin from H. sylvestris has been purified and functionally characterized, and two putative haemadins have been identified in the salivary transcriptome of Haemadipsa interrupta Moore, 1835, a terrestrial leech inhabiting mainly the Malayan peninsula. Here, we describe the identification of ten additional putative haemadins in the transcriptomic data set of H. interrupta, first generated by another study. Furthermore, we identified a putative oligomeric decorsin, which represents the first finding of this anticoagulant in a haemadipsid leech. Both the putative decorsin and a selection of haemadins were expressed, purified, and functionally characterized. The putative haemadins displayed a broad spectrum of thrombin-inhibitory potencies, whereas the putative oligomeric decorsin was indeed a weak inhibitor of platelet aggregation.

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Diversity of hirudin and hirudin-like factor genes in the North-African medicinal leech, Hirudo troctina

Ahmed,

Abilov,

Müller

2024

Parasitol Res

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Medicinal leeches of the genus Hirudo inhabit large areas of the Palaearctic realm. The distribution range of Hirudo troctina includes the southern Iberian peninsula and the northwestern regions of Africa. H. troctina is used for medical purposes, but only very little is known about the components of its salivary gland secretion. Hirudins, bivalent inhibitors of thrombin, are probably the best known leech-derived bioactive factors. Hirudin-like factors (HLFs) represent another class of salivary gland components that share characteristic genetic and structural markers with hirudins. Hirudin is not a single entity but exists in at least four different variants. However, there are differences among the European members of the genus Hirudo with respect to the actual number of hirudin and HLF genes that are present within their genomes. Here, we describe the identification and molecular cloning of 11 genes that encode for putative hirudin and HLF variants in H. troctina. Three of the genes consist of exons and introns that originate from different “archetype” genes and are likely the result of recombination events. The diversity of hirudin and HLF genes in H. troctina surpasses that of all other European members of the genus Hirudo. The putative hirudin variants and representatives of the HLFs of H. troctina were expressed as recombinant proteins, purified and functionally characterized for their thrombin-inhibiting potencies. Phylogenetic analyses based on hirudin and HLF gene sequences of the leech genera Hirudo, Hirudinaria, and Whitmania supported the hypothesis that hirudins and HLFs diverged early in leech evolution.

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The hirudin-like factors HLF3 and HLF4—hidden hirudins of European medicinal leeches

Cited by 12 publications

References 28 publications

Short tail stories: the hirudin-like factors HLF6 and HLF7 of the Asian medicinal leech, Hirudinaria manillensis

Short tail stories: the hirudin-like factors HLF6 and HLF7 of the Asian medicinal leech, Hirudinaria manillensis

Identification and functional characterization of multiple haemadins and an oligomeric decorsin in the Asian land leech Haemadipsa interrupta

Diversity of hirudin and hirudin-like factor genes in the North-African medicinal leech, Hirudo troctina

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