Isolation of a Fragment, Fh, Corresponding to the Hinge Region of Human IgG3

Michaelsen, T. E.; Natvig, J. B.; Sletten, K

doi:10.1111/j.1365-3083.1974.tb01282.x

Cited by 12 publications

(15 citation statements)

References 13 publications

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“…These findings suggest that there may be an extended hinge region in this subclass (7)(8)(9)(10). The isolation and determination of the sequence of this extra piece from the intact y3 chain have proven to be formidable projects, and to date, only the molecular weight and amino-acid composition have been reported (31). However, the availability of an unusual y3 HCD protein Omm-s, whose hinge appears to be identical to that of the normal y3 chain, has permitted the ready isolation of the extended hinge (Fh fragment) by simple chemical techniques.…”

Section: Discussionmentioning

confidence: 99%

Repetitive hinge region sequences in human IgG3: isolation of an 11,000-dalton fragment.

Adlersberg¹,

Frangione²

1975

Proc. Natl. Acad. Sci. U.S.A.

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The heavy chain (-y3) of the IgG3 subclass of human immunoglobulins has a molecular weight of 60,000, instead of the 50,000 value reported for -yl, y2, and The heavy chains of IgG (y) consist of four homology regions or "domains," each of which is made up of about 110 aminoacid residues, and is characterized by a single, highly conserved intrachain disulfide bridge (1). The amino-terminal domain, also known as the variable region (VH), participates in the formation of the antibody-combining site. The constant region, containing three domains (CH1, CH2, and CH3), is identical for molecules belonging to a given subclass and has approximately 90% homology from one y chain subclass to another (2, 3). Between the CH1 and CH2 domains, in the center of the 7 chain, is a region of unique sequence known as the "hinge" or interdomain, which has no homology with any known H or L chain domain. In the different subclasses of y chains, it contains a large number of proline residues, and a varying number of cysteine residues involved in the disulfide bridges linking the two heavy chains (4,5).Previous studies of a 73 myeloma protein (Kup) demonstrated the presence of five cysteine residues in a 33-residue Abbreviations: Nomenclature of immunoglobulins, their chains, and fragments follows the recommendations of the World Health Organization [WHO Bull. (1965) 33, 721; (1966) 35, 953; 38, 151; (1969)

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Section: Discussionmentioning

confidence: 99%

Repetitive hinge region sequences in human IgG3: isolation of an 11,000-dalton fragment.

Adlersberg¹,

Frangione²

1975

Proc. Natl. Acad. Sci. U.S.A.

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“…However, the hinge region of IgG1 and IgG3 differs greatly, first recognized in the early 70 ties [27][28][29] and later verified as hinge length of 15 aa for IgG1 and 62 aa for IgG3 respectively [30,31]. Intact hinge region, or at least one S-S bond at this site is necessary for complement activation [18,32].…”

Section: Discussion Hinge Differencesmentioning

confidence: 99%

Structural Difference in the Complement Activation Site of Human IgG1 and IgG3

et al. 2009

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The C1q binding epicentre on IgG molecules involves residues Asp270, Lys322, Pro329 and Pro331 in the CH2 domain. IgG1 and IgG3 are usually the most efficient of the four human IgG subclasses in activating complement and they both share all these residues. To reveal possible differences in the structural requirement for complement activation, we created a number of NIP (5‐iodo‐4‐hydroxy‐3‐nitro‐phenacetyl) specific IgG1 and IgG3 antibodies with parallel mutations in or near the putative C1q binding site. The mutants were tested simultaneously for antibody induced, antibody‐dependent complement‐mediated lysis (ADCML) at high and low antigen concentration on the target cells using sera of human, rabbit and guinea pig as complement source. In addition, we tested the antibodies against target cells decorated with the NP hapten, which has 10‐fold lower affinity for the antibodies compared to the NIP hapten. We also used ELISA methods to measure complement activation. We observed a clear difference between IgG1 and IgG3 localized to residues Asp270, Leu334, Leu335. For all these residues, and especially for Asp270, IgG1 was heavily reduced in complement activation, while IgG3 was only moderated reduced, by alanine substitution. This difference was independent of the long hinge region of IgG3, demonstrated by hinge region truncation of this isotype such that it resembles that of IgG1. This report indicates the presence of structural differences between human IgG1 and IgG3 in the C1q binding site, and points to a specialization of the two isotypes with respect to complement activation.

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“…Antisera, immunological quantitation, and dotible itnmttnodijfusion attalysis. The antisera were raised in rabbits and absorbed as previously described (11,13,14). Quantitation was performed by radial immunodiffusion analysis according to the method of Mancinl et al (8), as reported earlier (14).…”

Section: Methodsmentioning

confidence: 99%

“…Surprisingly, although papain is an endopeptidase with a broad specificit)', it splits at a limited numlicr of places in the hinge region and at only one peptide bond in the IgGl subclass (3). It has previously been reported that the IgG3 molecules have an extended hinge with an unusually open structure (10)(11)(12)(13)(14). It is therefore not surprising that the papain splits at several points in the hinge region of IgG3, resulting in two kinds of Fc fragments: Fch, whose hinge region to a large extent is intact, and Fc, which iacks most of the hinge region.…”

mentioning

confidence: 99%

Comparison of Fch and Fc Fragments of IgG3: Evidence of an Intra‐Chain S‐S Bridge in the Hinge of γ3 Chains

Michaelsen¹,

Natvig²

1974

Scand J Immunol

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The Fc and Fch fragments of human IgG3, appearing late and early during papain digestion, respectively, were antigenically and structurally compared. Only the Fch fragment reacted with an anti‐Fh (hinge region‐specific) antiserum; thus the main portion of the hinge region is split off during the formation of Fc fragments The Fch fragment has a higher content of inter‐chain S‐S bridges than the Fc fragments. The Fc fragments are probably a mixture of three related fragments. The Fch fragment appears to be more homogeneous, although there was hydrolysis at the C‐terminal site of the hinge region in some of the Fch chains. Evidence is presented that the γ3 chain bridge has an extra intra‐heavy‐chain disulphidc bridge at the C‐terminal end of the hinge region.

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Isolation of a Fragment, Fh, Corresponding to the Hinge Region of Human IgG3

Cited by 12 publications

References 13 publications

Repetitive hinge region sequences in human IgG3: isolation of an 11,000-dalton fragment.

Repetitive hinge region sequences in human IgG3: isolation of an 11,000-dalton fragment.

Structural Difference in the Complement Activation Site of Human IgG1 and IgG3

Comparison of Fch and Fc Fragments of IgG3: Evidence of an Intra‐Chain S‐S Bridge in the Hinge of γ3 Chains

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