Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reexamination Certificate
1999-10-05
2002-03-12
Eyler, Yvonne (Department: 1646)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S023500, C435S069100, C435S069700, C530S320000, C530S324000, C514S002600
Reexamination Certificate
active
06355788
ABSTRACT:
BACKGROUND OF THE INVENTION
Follistatin is a monomeric, glycosylated protein originally identified in porcine follicular fluid as a potent inhibitor of pituitary follicle-stimulating hormone (FSH) synthesis and secretion, follistatin was later shown to exert some of its biological effects by specifically binding the FSH-inducer activin. Other follistatin family members include follistatin related protein or FRP (Zwijsen et al.,
Eur. J. Biochem.
225:937-46, 1994), SPARC, also known as osteonectin or BM-40 or the human ortholog of mouse TSC-36 (Lane and Sage, ibid.), agrin (Patthy and Nikolics,
TINS
16:76-81, 1993), hevin (Girard and Springer,
Immunity
2:113-23, 1995), the Flik protein of chickens (Amthor et al.,
Dev. Biology
178:343-62, 1996) and the rat brain protein SC1 (Mendis et al.,
Brain Res.
730:95-106, 1996). Follistatins, however, are thought to be more than “activin binders” since follistatin deficient mice prepared by gene targeting have a more complex and different phenotype than activin gene knock-out animals (Mazuk et al.,
Nature
374:360-3, 1995 and Mazuk et al.,
Nature
374:356-9, 1995).
Activins and inhibins are potent activators and inhibitors, respectively, of pituitary FSH secretion and are members of the TGF-&bgr; family of peptide growth factors (Mather et al.,
Proc. Soc. Exp. Biol. Med.
215:209-22, 1997). The activin and inhibin family of hormones, while originally described as gonadally produced regulators of pituitary FSH secretion, are now known to have a broad range of effects within and outside of the reproductive system (Mather et al., ibid.). Inhibins consist of a common alpha subunit which is covalently linked to one of two different beta subunits (inhibin A:&agr;/B
A
; inhibin B:&agr;/B
B
); activins are covalently linked dimers of the two B-subunits and therefore exist in three different forms (activin A:B
A
/B
A
; activin B:B
B
/B
B
; activin AB:B
A
/B
B
). Activin and inhibin bind to follistatin with high affinity, and although the structure of the activin binding site has not been completely defined, preliminary data (Inouye et al.,
Biochem. Biophys. Res. Commun.
179:352-8, 1991) suggest that residues in the first amino terminal cysteine-rich follistatin domain are involved in hormone binding. Activin binding to follistatin is thus thought to limit its biological effects by sequestration of the peptide hormone. Thus, the broad range of biological actions of the activins and inhibins, and possibly other members of the TGF-&bgr; family as well, may be regulated by binding to proteins of the follistatin family. Different binding proteins may be involved for each TGF-&bgr; family member as follistatin binds activin with high affinity (nM), inhibin with lower affinity, and does not appear to bind TGF-&bgr; at all (Mather et al., ibid.). Follistatin family members may regulate the activity of other growth factors as well, for example, SPARC or BM-40 have been shown to bind platelet derived growth factor (PDGF-AB, PDGF-BB) (Lane and Sage,
FASEB J.
8:163-73, 1994).
This application provides a new member of the follistatin family, zfsta2, which is likely to play a major role in regulating the biological activities of the TGF-&bgr; growth factors. Like other members of the follistatin family, zfsta2 may play a broad role in development and differentiation, pathogenesis of atherosclerosis, regulation of the gonadal-pituitary-hypothalamic axis, tooth and bone formation, regulation of gonadal hormone production, spermatogenesis, hypothalmic oxytocin secretion, proliferation and differentiation of erythroid progenitors, hematopoiesis, host defense and neuron survival.
The present invention provides such polypeptides for these and other uses that should be apparent to those skilled in the art from the teachings herein.
SUMMARY OF THE INVENTION
Within one aspect the invention provides an isolated polypeptide comprising a follistatin homology domain, wherein the follistatin homology domain comprises amino acid residues 65 to 133 of the amino acid sequence of SEQ ID NO:2. Within one embodiment the polypeptide further comprises an alpha helical linker region that resides in a carboxyl-terminal position relative to the follistatin homology domain, wherein the alpha helical linker region comprises amino acid residues 134 to 174 of the amino acid sequence of SEQ ID NO:2. Within a related embodiment the polypeptide further comprises a calmodulin homology domain that resides in a carboxyl-terminal position relative to the alpha helical linker region, wherein the calmodulin homology domain comprises amino acid residues 175 to 250 of the amino acid sequence of SEQ ID NO:2. Within another embodiment the polypeptide further comprises two I-set Ig domains that reside in a carboxyl-terminal position relative to the calmodulin homology domain, wherein the I-set Ig domains comprise amino acid residues 251 to 431 of the amino acid sequence of SEQ ID NO:2. Within another embodiment the polypeptide further comprises a carboxy-terminal domain that resides in a carboxyl-terminal position relative to the I-set Ig domains, wherein the carboxy-terminal domain comprises amino acid residues 433 to 847 of the amino acid sequence of SEQ ID NO:2. Within a yet another embodiment the polypeptide further comprises a hydrophilic linker region that resides in an amino-terminal position relative to the follistatin homology domain, wherein the hydrophobic linker region comprises amino acid residues 21 to 64 of the amino acid sequence of SEQ ID NO:2. Within another embodiment the polypeptide further comprises a secretory signal sequence that resides in an amino-terminal position relative to the hydrophobic linker region, wherein the secretory signal sequence comprises amino acid residues 1 to 20 of the amino acid sequence of SEQ ID NO:2.
Within another aspect the invention provides an isolated polypeptide having an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO:2, wherein the isolated polypeptide specifically binds with an antibody to which a polypeptide having the amino acid sequence of SEQ ID NO:2 specifically binds. Within one embodiment the isolated polypeptide has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:2. Within another embodiment the isolated polypeptide has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2. Within a further embodiment any difference between the amino acid sequence and the corresponding amino acid sequence of SEQ ID NO:2 is due to one or more conservative amino acid substitutions. Within another embodiment the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=blosum62, with other parameters set as default.
The invention also provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO:2.
The invention further provides an isolated polypeptide selected from the group consisting of: a) a polypeptide consisting of the sequence of amino acid residues from residue 21 to residue 64 of SEQ ID NO:2; b) a polypeptide consisting of the sequence of amino acid residues from residue 65 to residue 133 of SEQ ID NO:2; c) a polypeptide consisting of the sequence of amino acid residues from residue 134 to residue 174 of SEQ ID NO:2; d) a polypeptide consisting of the sequence of amino acid residues from residue 175 to residue 250 of SEQ ID NO:2; e) a polypeptide consisting of the sequence of amino acid residues from residue 251 to residue 334 of SEQ ID NO:2; f) a polypeptide consisting of the sequence of amino acid residues from residue 335 to residue 432 of SEQ ID NO:2; g) a polypeptide consisting of the sequence of amino acid residues from residue 433 to residue 847 of SEQ ID NO:2; h) a polypeptide consisting of the sequence of amino acid residues from residue 251 to residue 432 of SEQ ID NO:2; i) a polypeptide consisting of the sequence of amino acid residues from residue 65 to res
Conklin Darrell C.
Ellsworth Jeff L.
Andres Janet L.
Eyler Yvonne
Lingenfelter Susan E.
ZymoGenetics Inc.
LandOfFree
Follistatin-related protein zfsta2 does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Follistatin-related protein zfsta2, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Follistatin-related protein zfsta2 will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2889291