Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues
Reexamination Certificate
2000-04-19
2003-09-16
McKelvey, Terry (Department: 1636)
Chemistry: natural resins or derivatives; peptides or proteins;
Peptides of 3 to 100 amino acid residues
C530S324000, C530S350000
Reexamination Certificate
active
06620909
ABSTRACT:
BACKGROUND OF THE INVENTION
Energy balance (involving energy metabolism, nutritional state, lipid storage and the like) is an important criteria for health. This energy homeostasis involves food intake and metabolism of carbohydrates and lipids to generate energy necessary for voluntary and involuntary functions. Metabolism of proteins can lead to energy generation, but preferably leads to muscle formation or repair. Among other consequences, a lack of energy homeostasis lead to over or under formation of adipose tissue.
Formation and storage of fat is insulin-modulated. For example, insulin stimulates the transport of glucose into cells, where it is metabolized into &agr;-glycerophosphate which is used in the esterification of fatty acids to permit storage thereof as triglycerides. In addition, adipocytes (fat cells) express a specific transport protein that enhances the transfer of free fatty acids into adipocytes.
Adipocytes also secrete several proteins believed to modulate homeostatic control of glucose and lipid metabolism. These additional adipocyte-secreted proteins include adipsin, complement factors C3 and B, tumor necrosis factor &agr;, the ob gene product and Acrp30. Evidence also exists suggesting the existence of an insulin-regulated secretory pathway in adipocytes. Scherer et al.,
J. Biol. Chem.
270(45): 26746-9, 1995. Over or under secretion of these moieties, impacted in part by over or under formation of adipose tissue, can lead to pathological conditions associated directly or indirectly with obesity or anorexia.
Acrp30 is a 247 amino acid polypeptide that is expressed exclusively by adipocytes. The Acrp30 polypeptide is composed of a amino-terminal signal sequence, a 27 amino acid stretch of no known homology, 22 perfect Gly-Xaa-Pro or imperfect Gly-Xaa-Xaa collagen repeats and a carboxy terminal globular domain. See, Scherer et al. as described above and International Patent Application No. WO 96/39429. Acrp30, an abundant human serum protein regulated by insulin, shares structural similarity, particularly in the carboxy-terminal globular domain, to complement factor Clq and to a summer serum protein of hibernating Siberian chipmunks (Hib27). Expression of Acrp30 is induced over 100-fold during adipocyte differentiation. Acrp30 is suggested for use in modulating energy balance and in identifying adipocytes in test samples.
Another secreted protein that appears to be exclusively produced in adipocytes is apM1, described, for example, in Maeda et al.,
Biochem. Biophys. Res. Comm.
221: 286-9, 1996. A 4517 bp clone had a 244 amino acid open reading frame and a long 3′ untranslated region. The protein included a signal sequence, an amino-terminal non-collagenous sequence, 22 collagen repeats (Gly-XAA-Pro or Gly-Xaa-Xaa), and a carboxy-terminal region with homology to collagen X, collagen VIII and complement protein Clq.
Complement factor Clq consists of six copies of three related polypeptides (A, B and C chains), with each polypeptide being about 225 amino acids long with a near amino-terminal collagen domain and a carboxy-terminal globular region. Six triple helical regions are formed by the collagen domains of the six A, six B and six C chains, forming a central region and six stalks. A globular head portion is formed by association of the globular carboxy terminal domain of an A, a B and a C chain. Clq is therefore composed of six globular heads linked via six collagen-like stalks to a central fibril region. Sellar et al.,
Biochem. J.
274: 481-90, 1991. This configuration is often referred to as a bouquet of flowers. Acrp30 has a similar bouquet structure formed from a single type of polypeptide chain.
Clq has been found to stimulate defense mechanisms as well as trigger the generation of toxic oxygen species that can cause tissue damage (Tenner,
Behring Inst. Mitt.
93:241-53, 1993). Clq binding sites are found on platelets. Additionally complement and Clq play a role in inflammation. The complement activation is initiated by binding of Clq to immunoglobulins.
Inhibitors of Clq and the complement pathway would be useful for anti-inflammatory applications, inhibition of complement activation and thrombotic activity.
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 sequence of amino acid residues that is at least 75% identical in amino acid sequence to residues 40-285 of SEQ ID NO:2, wherein the sequence comprises: Gly-Xaa-Xaa or Gly-Xaa-Pro repeats forming a collagen domain, wherein Xaa is any amino acid; and a carboxyl-terminal Clq domain comprises 10 beta strands. Within one embodiment the polypeptide that is at least 90% identical in amino acid sequence to residues 16-285 of SEQ ID NO:2. Within another embodiment the collagen domain consists of 24 Gly-Xaa-Xaa repeats and 10 Gly-Xaa-Pro repeats. Within another embodiment the carboxyl-terminal Clq domain comprises the sequence of SEQ ID NO:5. Within another embodiment the carboxy-terminal Clq domain comprises amino acid residues 151-155, 172-174, 180-183, 187-190, 193-205, 208-214, 220-227, 229-241, 246-251 and 269-274 of SEQ ID NO:2. Within another embodiment any differences between said polypeptide and SEQ ID NO:2 are due to conservative amino acid substitutions. Within another embodiment the polypeptide specifically binds with an antibody that specifically binds with a polypeptide consisting of the amino acid sequence of SEQ ID NO:2. Within a further embodiment the polypeptide comprises residues 16-285 of SEQ ID NO:2. Within another embodiment the polypeptide is covalently linked at the amino or carboxyl terminus to a moiety selected from the group consisting of affinity tags, toxins, radionucleotides, enzymes and fluorophores. Within yet another embodiment the collagen domain consists of amino acid residues 41-141 of SEQ ID NO:2. Within another embodiment the carboxy-terminal Clq domain consists of amino acid residues 142-285 of SEQ ID NO:2.
The invention also provides an isolated polypeptide selected from the group consisting of: a) a polypeptide consisting of a sequence of amino acid residues that is 75% identical in amino acid sequence to amino acid residue 40 to amino acid residue 141 of SEQ ID NO:2; b) a polypeptide consisting of a sequence of amino acid residues that is 75% identical in amino acid sequence to amino acid residue 142 to amino acid residue 285 of SEQ ID NO:2; and c) a polypeptide consisting of a sequence of amino acid residues that is 75% identical in amino acid sequence to amino acid residue 40 to 285 of SEQ ID NO:2.
Within another aspect, the invention provides a fusion protein comprising a first portion and a second portion joined by a peptide bond, the first portion consisting of a polypeptide selected from the group consisting of: a) a polypeptide comprising a sequence of amino acid residues that is at least 75% identical in amino acid sequence to amino acid residue 16 to amino acid residue 285 of SEQ ID NO:2; b) a polypeptide comprising a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid residue 1 to amino acid residue 281; c) a polypeptide comprising a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid residue 16 to amino acid residue 285; d) a portion of the zacrp2 polypeptide as shown in SEQ ID NO:2, comprising the collagen-like domain or a portion of the collagen-like domain capable of dimerization or oligomerization; e) a portion of the zacrp2 polypeptide as shown in SEQ ID NO:2, comprising the Clq domain or an active portion of the Clq domain; or f) a portion of the zacrp2 polypeptide as shown in SEQ ID NO:2 comprising of the collagen-like domain and the Clq domain; and the second portion comprising another polypeptide. Within one embodiment the first portion is selected from the group consisting of: a) a polypeptide consisting of the sequence of amino acid residue 40 to amino acid residue 141 of SEQ ID NO:2; b)
Bishop Paul D.
Piddington Christopher S.
McKelvey Terry
Walsh Brian J.
ZymoGenetics Inc.
LandOfFree
Adipocyte complement related protein homolog zacrp2 does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Adipocyte complement related protein homolog zacrp2, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Adipocyte complement related protein homolog zacrp2 will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3004656